We examined, in vitro, whether hyaluronan induces slow cycling in placenta-derived mesenchymal st... more We examined, in vitro, whether hyaluronan induces slow cycling in placenta-derived mesenchymal stem cells (PDMSCs) by comparing cell growth on a hyaluronan-coated surface with cell growth on a tissueculture polystyrene surface. The hyaluronan-coated surface significantly downregulated the proliferation of PDMSCs, more of which were maintained in the G 0 /G 1 phases than were cells on the tissue-culture polystyrene surface. Both PKH-26 labeling and BrdU incorporation assays showed that most PDMSCs grown on a hyaluronan-coated surface duplicated during cultivation indicating that the hyaluronan-coated surface did not inhibit PDMSCs from entering the cell cycle. Mitotic synchronization showed that the G 1-phase transit was prolonged in PDMSCs growing on a hyaluronan-coated surface. Increases in p27 Kip1 and p130 were the crucial factors that allowed hyaluronan to lengthen the G 1 phase. Thus, hyaluronan might be a promising candidate for maintaining stem cells in slow-cycling mode by prolonging their G 1-phase transit.
Journal of Biomaterials Science-polymer Edition, 1999
Bioadhesives have been used in surgery as hemostatic and wound healing agents. GRF (gelatin + res... more Bioadhesives have been used in surgery as hemostatic and wound healing agents. GRF (gelatin + resorcinol + formaldehyde) glue, composed of a mixture of gelatin and resorcinol polymerized by the addition of formaldehyde, has been used for this purpose. Widespread acceptance of the GRF glue, however, has been limited by reports of cytotoxicity due to its release of formaldehyde upon degradation. It has been suggested by Wertzel et al. that the cytotoxicity problem of GRF glue may be overcome by changing its cross-linking method. The study was, therefore, undertaken to assess the feasibility of using a water-soluble carbodiimide or genipin to cross-link gelatin as new bioadhesives to close skin wound lesions in a rat model. Formaldehyde-cross-linked counterpart (GRF glue) and a resorbable suture were used as controls. It was noted that the tensile strength of the skin across each wound treated by either application of test glues or suture increased consistently with time during the healing process. Also, the wounds repaired by test glues or suture caused no calcification. The suture used in the study was completely resorbed at the wound area in about 6 days postoperatively. However, the durations required to completely resorb the carbodiimide- or genipin-cross-linked glues were approximately the same (9 days), while it took about 14 days to completely resorb the formaldehyde-cross-linked glue. The healing process for the suture wound repaired was more rapid than those treated by test glues. Of the test glues, the wounds treated by the carbodiimide- or genipin-cross-linked glues induced less inflammatory response and recovered sooner than that treated by the formaldehyde-cross-linked glue. This indicated that the biocompatibility of the carbodiimide- or genipin-cross-linked glues was superior to the formaldehyde-cross-linked glue. The results of this study may serve as a preliminary experimental model for the further investigation of both the carbodiimide- and genipin-cross-linked glues when applied to human skin closure.
Based on plastically compressed cell-seeded collagen gels, we fabricated a small-diameter tubular... more Based on plastically compressed cell-seeded collagen gels, we fabricated a small-diameter tubular construct that withstands arterial pressure without prolonged culture in vitro. Specifically, to mimic the microstructure of vascular media, the cell-seeded collagen gel was uniaxially stretched prior to plastic compression to align collagen fibers and hence cells in the gel. The resulting gel sheet was then wrapped around a custom-made multi-layered braided tube to form aligned tubular constructs whereas the gel sheet prepared similarly but without uniaxial stretching formed control constructs. With the braided tube, fluid in the gel construct was further removed by vacuum suction aiming to consolidate the concentric layers of the construct. The construct was finally treated with transglutaminase. Both SEM and histology confirmed the absence of gaps in the wall of the construct. Particularly, cells in the wall of the aligned tubular construct were circumferentially aligned. The enzyme-mediated crosslinking increased burst pressure of both the constructs significantly; the extent of the increase of burst pressure for the aligned tubular construct was greater than that for the control counterpart. Increasing crosslinking left the compliance of the aligned tubular construct unchanged but reduced that of the control construct. Cells remained viable in transglutaminase-treated plastically compressed gels after 6 days in culture. This study demonstrated that by combining stretch-induced fiber alignment, plastic compression, and enzyme-mediated crosslinking, a cell-seeded collagen gel-based tubular construct with potential to be used as vascular media can be made within 3 days.
Glioblastoma multiforme (GBM) is one of the most malignant and aggressive brain tumors with great... more Glioblastoma multiforme (GBM) is one of the most malignant and aggressive brain tumors with great amount of hyaluronan (HA) secretion and CD44 overexpression (HA receptor). CD44 has been suggested as a cancer stem cells (CSCs) marker. However, several clinical studies have indicated that CD44low glioma cell exhibit CSCs traits. Additionally, our previous study indicated that more CD44 expression was associated with a better prognosis in GBM patients. To determine whether CD44 is an appropriate marker of glioma stem cells (GSCs), we manipulated CD44 expression using intrinsic (CD44 knockdown, CD44kd) and extrinsic (HA supplement, HA+) methods. Our results show that CD44kd suppressed cell proliferation by retarding cell cycle progression from G0/G1 to S phase. Furthermore, it caused GSCs traits, including lower expression of differentiation marker (glial fibrillary acidic protein, GFAP), a higher level of sphere formation and higher expression of stem cell markers (CD133, nestin and Oct4). The reduction of CD44 expression induced by HA+ was accompanied by an increase in GSCs properties. Interestingly, the presence of HA+ in glioma cells with GSC traits conversely facilitated differentiation. Our data indicated that the CD44 low-expressing cells exhibit more GSCs straits, suggesting that CD44 is not an appropriate marker for GSCs. Furthermore, the preferential expression of CD44 at the invasive rim in rat glioma specimen implies that CD44 may be more important for invasion and migration instead of GSCs marker in glioma.
In this study, the intensity of two-photon excited fluorescence (TPEF) of xanthene dye, Rose Beng... more In this study, the intensity of two-photon excited fluorescence (TPEF) of xanthene dye, Rose Bengal (RB), was significantly enhanced via bovine serum albumin (BSA) microstructures fabricated by the two-photon crosslinking (TPC) technique. The RB was utilized as the photoactivator in the TPC processing and the enhanced TPEF intensity correlates with the concentration of fabricated crosslinked BSA microstructures via the power control and pulse selection of the employed femtosecond laser. As a result, fabrication of three-dimensional BSA microstructures can be simultaneously monitored by the use of TPEF intensity. The crosslinked BSA microstructures synthesized may be used as an ordered biomaterial for fluorescence enhancement.
Hyaluronan (HA), an abundant polysaccharide found in human bodies, plays a role in the mesenchyma... more Hyaluronan (HA), an abundant polysaccharide found in human bodies, plays a role in the mesenchymal stem cells (MSCs) maintenance. We had previously found that HA prolonged the lifespan, and prevented the cellular aging of murine adiposederived stromal cells. Recently, we had also summarized the potential pathways associated with HA regulation in human MSCs. In this study, we used the human placenta-derived MSCs (PDMSC) to investigate the effectiveness of HA in maintaining the PDMSC. We found that coating the culture surface coated with 30 lg cm À2 of HA (C) led to cluster growth of PDMSC, and maintained a higher number of PDMSC in quiescence compared to those grown on the normal tissue culture surface (T). PDMSC were treated for either 4 (short-term) or 19 (long-term) consecutive passages. PDMSC which were treated with HA for 19 consecutive passages had reduced cell enlargement, preserved MSCs biomarker expressions and osteogenic potential when compared to those grown only on T. The PDMSC transferred to T condition after long-term HA treatment showed preserved replicative capability compared to those on only T. The telomerase activity of the HA-treated PDMSC was also higher than that of untreated PDMSC. These data suggested a connection between HA and MSC maintenance. We suggest that HA might be regulating the distribution of cytoskeletal proteins on cell spreading in the event of quiescence to preserve MSC stemness. Maintenance of MSCs stemness delayed cellular aging, leading to the anti-aging phenotype of PDMSC.
An appropriate animal wound model is urgently needed to assess wound dressings, cell therapies, a... more An appropriate animal wound model is urgently needed to assess wound dressings, cell therapies, and pharmaceutical agents. Minipig was selected owing to similarities with humans in body size, weight, and physiological status. Different wound sizes (0.07-100 cm 2) were created at varying distances but fail to adequately distinguish the efficacy of various interventions. We aimed to resolve potential drawbacks by developing a systematic wound healing system. No significant variations in dorsal wound closure and contraction were observed within the thoracolumbar region between boundaries of both armpits and the paravertebral region above rib tips; therefore, Lanyu pigs appear suitable for constructing a reliable dorsal wound array. Blood flow signals interfered with inter-wound distances ˂ 4 cm; a distance > 4 cm is therefore recommended. Wound sizes ≥ 4 cm × 4 cm allowed optimal differentiation of interventions. Partial-(0.23 cm) and full-thickness (0.6 cm) wounds showed complete re-epithelialization on days 13 and 18 and strongest blood flow signals at days 4 and 11, respectively. Given histological and tensile strength assessments, tissue healing resembling normal skin was observed at least after 6 months. We established some golden standards for minimum wound size and distance between adjacent wounds for effectively differentiating interventions in considering 3R principles. An ideal preclinical animal model is crucial for assessing wound healing and management to meet the growing demand for developing biomedical products 1,2. Animal models simulating distinct human skin wound conditions have been developed to evaluate the effectiveness of different interventions 3-8. Nevertheless, some drawbacks persist in existing models 9. The loosened skin of small laboratory mammals, including rodents and rabbits, differs from human skin, closing the wounds via contraction. This characteristic, on the contrary, may benefit for evaluation of anti-contraction effects of biomaterials in wounds. Small animal selection is based on considerations such as laboratory space and cost 4-8 ; however, these animals afford limited skin surface area, necessitating numerous animals to acquire sufficient data 10,11 , leading to errors and inconsistencies in the analyses of results. Although sheep, goats, dogs, and other animals were also selected for external traumatic wound studies, they are more suitable for studying aspects such as bone, ligaments, and tendons. In contrast, pigs are better for wound studies due to larger dorsal surface area. Recently, pigs have been widely employed as a model for biomedical investigations because of their tight skin system that resembles that of humans, with a thick epidermis, distinct rete pegs, dense collagen fibrils, elastic fibrils in the dermis, and accessory hair and sweat glands. Both humans and pigs also demonstrate an identical wound healing mechanism, including restoration of the dermis, skin re-epithelialization, and skin contraction 12-14. Furthermore, the body size of pigs is sufficiently large to provide an adequate skin surface to create multiple wounds on the same animal, thereby improving the reliability of results by eliminating extraneous factors associated with different animals 15,16. In the last decade, commercially bred pig varieties such as Duroc, Yorkshire, and Landrace domestic pigs 15-17 have been employed in skin wound healing studies. Despite the clinical validity and applicability of porcine models, larger breeding spaces and
Advances in Experimental Medicine and Biology, 2018
In order to develop and commercialize for the regenerative medicinal products, smart biomaterials... more In order to develop and commercialize for the regenerative medicinal products, smart biomaterials with biocompatibility must be needed. In this chapter, we introduce collagen and hyaluronic acid (HA) as extracellular matrix as well as deal with the molecular mechanism as microenvironment, mechanistic effects, and gene expression. Application of collagen and HA have been reviewed in the area of orthopedics, orthopedics, ophthalmology, dermatology and plastic surgery. Finally, the ongoing and commercial products of collagen and HA for regenerative medicine have been introduced.
Acanthamoeba is free-living protist pathogen capable of causing a blinding keratitis and granulom... more Acanthamoeba is free-living protist pathogen capable of causing a blinding keratitis and granulomatous encephalitis. However, the mechanisms of Acanthamoeba pathogenesis are still not clear. Here, our results show that cells co-cultured with pathogenic Acanthamoeba would be spherical and floated, even without contacting the protists. Then, the Acanthamoeba protists would contact and engulf these cells. In order to clarify the contact-independent pathogenesis mechanism in Acanthamoeba, we collected the Acanthamoeba-secreted proteins (Asp) to incubate with cells for identifying the extracellular virulent factors and investigating the cytotoxicity process. The Asps of pathogenic Acanthamoeba express protease activity to reactive Leu amino acid in ECM and induce cell-losing adhesion ability. The M20/M25/M40 superfamily aminopeptidase protein (ACA1_264610), an aminopeptidase be found in Asp, is upregulated after Acanthamoeba and C6 cell co-culturing for 6 h. Pre-treating the Asp with leucine aminopeptidase inhibitor and the specific antibodies of Acanthamoeba M20/M25/M40 superfamily aminopeptidase could reduce the cell damage during Asp and cell co-incubation. These results suggest an important functional role of the Acanthamoeba secreted extracellular aminopeptidases in the Acanthamoeba pathogenesis process. This study provides information regarding clinically pathogenic isolates to target specific molecules and design combined drugs.
This study has demonstrated that baicalein has anticancer effectiveness against human hepatoma ce... more This study has demonstrated that baicalein has anticancer effectiveness against human hepatoma cells. The dose response of baicalein in Hep G2 and Hep J2 cells indicates that baicalein decreased viability >90%. In comparison, baicalein had only minimal effects on the viability of control Chang liver cells. Flow cytometric analysis showed that baicalein inhibited the cell cycle of Hep G2 cells in the S phase. In addition, baicalein treatment resulted in a decreased mitochondrial transmembrane potential and damaged the integrity of the cell membrane. The TdT-mediated dUTP-biotin nick end labeling assay results indicated that baicalein elicited a significant increase of DNA fragmentation in Hep G2 cells after incubation for 48 hours. These results indicate that baicalein is an effective antihepatoma agent with minimal influence on noncancer cells. The effects of baicalein on Hep G2 cells include inhibition of the S phase of the cell cycle, dysfunction of mitochondria, and initiation of apoptosis.
We examined, in vitro, whether hyaluronan induces slow cycling in placenta-derived mesenchymal st... more We examined, in vitro, whether hyaluronan induces slow cycling in placenta-derived mesenchymal stem cells (PDMSCs) by comparing cell growth on a hyaluronan-coated surface with cell growth on a tissueculture polystyrene surface. The hyaluronan-coated surface significantly downregulated the proliferation of PDMSCs, more of which were maintained in the G 0 /G 1 phases than were cells on the tissue-culture polystyrene surface. Both PKH-26 labeling and BrdU incorporation assays showed that most PDMSCs grown on a hyaluronan-coated surface duplicated during cultivation indicating that the hyaluronan-coated surface did not inhibit PDMSCs from entering the cell cycle. Mitotic synchronization showed that the G 1-phase transit was prolonged in PDMSCs growing on a hyaluronan-coated surface. Increases in p27 Kip1 and p130 were the crucial factors that allowed hyaluronan to lengthen the G 1 phase. Thus, hyaluronan might be a promising candidate for maintaining stem cells in slow-cycling mode by prolonging their G 1-phase transit.
Journal of Biomaterials Science-polymer Edition, 1999
Bioadhesives have been used in surgery as hemostatic and wound healing agents. GRF (gelatin + res... more Bioadhesives have been used in surgery as hemostatic and wound healing agents. GRF (gelatin + resorcinol + formaldehyde) glue, composed of a mixture of gelatin and resorcinol polymerized by the addition of formaldehyde, has been used for this purpose. Widespread acceptance of the GRF glue, however, has been limited by reports of cytotoxicity due to its release of formaldehyde upon degradation. It has been suggested by Wertzel et al. that the cytotoxicity problem of GRF glue may be overcome by changing its cross-linking method. The study was, therefore, undertaken to assess the feasibility of using a water-soluble carbodiimide or genipin to cross-link gelatin as new bioadhesives to close skin wound lesions in a rat model. Formaldehyde-cross-linked counterpart (GRF glue) and a resorbable suture were used as controls. It was noted that the tensile strength of the skin across each wound treated by either application of test glues or suture increased consistently with time during the healing process. Also, the wounds repaired by test glues or suture caused no calcification. The suture used in the study was completely resorbed at the wound area in about 6 days postoperatively. However, the durations required to completely resorb the carbodiimide- or genipin-cross-linked glues were approximately the same (9 days), while it took about 14 days to completely resorb the formaldehyde-cross-linked glue. The healing process for the suture wound repaired was more rapid than those treated by test glues. Of the test glues, the wounds treated by the carbodiimide- or genipin-cross-linked glues induced less inflammatory response and recovered sooner than that treated by the formaldehyde-cross-linked glue. This indicated that the biocompatibility of the carbodiimide- or genipin-cross-linked glues was superior to the formaldehyde-cross-linked glue. The results of this study may serve as a preliminary experimental model for the further investigation of both the carbodiimide- and genipin-cross-linked glues when applied to human skin closure.
Based on plastically compressed cell-seeded collagen gels, we fabricated a small-diameter tubular... more Based on plastically compressed cell-seeded collagen gels, we fabricated a small-diameter tubular construct that withstands arterial pressure without prolonged culture in vitro. Specifically, to mimic the microstructure of vascular media, the cell-seeded collagen gel was uniaxially stretched prior to plastic compression to align collagen fibers and hence cells in the gel. The resulting gel sheet was then wrapped around a custom-made multi-layered braided tube to form aligned tubular constructs whereas the gel sheet prepared similarly but without uniaxial stretching formed control constructs. With the braided tube, fluid in the gel construct was further removed by vacuum suction aiming to consolidate the concentric layers of the construct. The construct was finally treated with transglutaminase. Both SEM and histology confirmed the absence of gaps in the wall of the construct. Particularly, cells in the wall of the aligned tubular construct were circumferentially aligned. The enzyme-mediated crosslinking increased burst pressure of both the constructs significantly; the extent of the increase of burst pressure for the aligned tubular construct was greater than that for the control counterpart. Increasing crosslinking left the compliance of the aligned tubular construct unchanged but reduced that of the control construct. Cells remained viable in transglutaminase-treated plastically compressed gels after 6 days in culture. This study demonstrated that by combining stretch-induced fiber alignment, plastic compression, and enzyme-mediated crosslinking, a cell-seeded collagen gel-based tubular construct with potential to be used as vascular media can be made within 3 days.
Glioblastoma multiforme (GBM) is one of the most malignant and aggressive brain tumors with great... more Glioblastoma multiforme (GBM) is one of the most malignant and aggressive brain tumors with great amount of hyaluronan (HA) secretion and CD44 overexpression (HA receptor). CD44 has been suggested as a cancer stem cells (CSCs) marker. However, several clinical studies have indicated that CD44low glioma cell exhibit CSCs traits. Additionally, our previous study indicated that more CD44 expression was associated with a better prognosis in GBM patients. To determine whether CD44 is an appropriate marker of glioma stem cells (GSCs), we manipulated CD44 expression using intrinsic (CD44 knockdown, CD44kd) and extrinsic (HA supplement, HA+) methods. Our results show that CD44kd suppressed cell proliferation by retarding cell cycle progression from G0/G1 to S phase. Furthermore, it caused GSCs traits, including lower expression of differentiation marker (glial fibrillary acidic protein, GFAP), a higher level of sphere formation and higher expression of stem cell markers (CD133, nestin and Oct4). The reduction of CD44 expression induced by HA+ was accompanied by an increase in GSCs properties. Interestingly, the presence of HA+ in glioma cells with GSC traits conversely facilitated differentiation. Our data indicated that the CD44 low-expressing cells exhibit more GSCs straits, suggesting that CD44 is not an appropriate marker for GSCs. Furthermore, the preferential expression of CD44 at the invasive rim in rat glioma specimen implies that CD44 may be more important for invasion and migration instead of GSCs marker in glioma.
In this study, the intensity of two-photon excited fluorescence (TPEF) of xanthene dye, Rose Beng... more In this study, the intensity of two-photon excited fluorescence (TPEF) of xanthene dye, Rose Bengal (RB), was significantly enhanced via bovine serum albumin (BSA) microstructures fabricated by the two-photon crosslinking (TPC) technique. The RB was utilized as the photoactivator in the TPC processing and the enhanced TPEF intensity correlates with the concentration of fabricated crosslinked BSA microstructures via the power control and pulse selection of the employed femtosecond laser. As a result, fabrication of three-dimensional BSA microstructures can be simultaneously monitored by the use of TPEF intensity. The crosslinked BSA microstructures synthesized may be used as an ordered biomaterial for fluorescence enhancement.
Hyaluronan (HA), an abundant polysaccharide found in human bodies, plays a role in the mesenchyma... more Hyaluronan (HA), an abundant polysaccharide found in human bodies, plays a role in the mesenchymal stem cells (MSCs) maintenance. We had previously found that HA prolonged the lifespan, and prevented the cellular aging of murine adiposederived stromal cells. Recently, we had also summarized the potential pathways associated with HA regulation in human MSCs. In this study, we used the human placenta-derived MSCs (PDMSC) to investigate the effectiveness of HA in maintaining the PDMSC. We found that coating the culture surface coated with 30 lg cm À2 of HA (C) led to cluster growth of PDMSC, and maintained a higher number of PDMSC in quiescence compared to those grown on the normal tissue culture surface (T). PDMSC were treated for either 4 (short-term) or 19 (long-term) consecutive passages. PDMSC which were treated with HA for 19 consecutive passages had reduced cell enlargement, preserved MSCs biomarker expressions and osteogenic potential when compared to those grown only on T. The PDMSC transferred to T condition after long-term HA treatment showed preserved replicative capability compared to those on only T. The telomerase activity of the HA-treated PDMSC was also higher than that of untreated PDMSC. These data suggested a connection between HA and MSC maintenance. We suggest that HA might be regulating the distribution of cytoskeletal proteins on cell spreading in the event of quiescence to preserve MSC stemness. Maintenance of MSCs stemness delayed cellular aging, leading to the anti-aging phenotype of PDMSC.
An appropriate animal wound model is urgently needed to assess wound dressings, cell therapies, a... more An appropriate animal wound model is urgently needed to assess wound dressings, cell therapies, and pharmaceutical agents. Minipig was selected owing to similarities with humans in body size, weight, and physiological status. Different wound sizes (0.07-100 cm 2) were created at varying distances but fail to adequately distinguish the efficacy of various interventions. We aimed to resolve potential drawbacks by developing a systematic wound healing system. No significant variations in dorsal wound closure and contraction were observed within the thoracolumbar region between boundaries of both armpits and the paravertebral region above rib tips; therefore, Lanyu pigs appear suitable for constructing a reliable dorsal wound array. Blood flow signals interfered with inter-wound distances ˂ 4 cm; a distance > 4 cm is therefore recommended. Wound sizes ≥ 4 cm × 4 cm allowed optimal differentiation of interventions. Partial-(0.23 cm) and full-thickness (0.6 cm) wounds showed complete re-epithelialization on days 13 and 18 and strongest blood flow signals at days 4 and 11, respectively. Given histological and tensile strength assessments, tissue healing resembling normal skin was observed at least after 6 months. We established some golden standards for minimum wound size and distance between adjacent wounds for effectively differentiating interventions in considering 3R principles. An ideal preclinical animal model is crucial for assessing wound healing and management to meet the growing demand for developing biomedical products 1,2. Animal models simulating distinct human skin wound conditions have been developed to evaluate the effectiveness of different interventions 3-8. Nevertheless, some drawbacks persist in existing models 9. The loosened skin of small laboratory mammals, including rodents and rabbits, differs from human skin, closing the wounds via contraction. This characteristic, on the contrary, may benefit for evaluation of anti-contraction effects of biomaterials in wounds. Small animal selection is based on considerations such as laboratory space and cost 4-8 ; however, these animals afford limited skin surface area, necessitating numerous animals to acquire sufficient data 10,11 , leading to errors and inconsistencies in the analyses of results. Although sheep, goats, dogs, and other animals were also selected for external traumatic wound studies, they are more suitable for studying aspects such as bone, ligaments, and tendons. In contrast, pigs are better for wound studies due to larger dorsal surface area. Recently, pigs have been widely employed as a model for biomedical investigations because of their tight skin system that resembles that of humans, with a thick epidermis, distinct rete pegs, dense collagen fibrils, elastic fibrils in the dermis, and accessory hair and sweat glands. Both humans and pigs also demonstrate an identical wound healing mechanism, including restoration of the dermis, skin re-epithelialization, and skin contraction 12-14. Furthermore, the body size of pigs is sufficiently large to provide an adequate skin surface to create multiple wounds on the same animal, thereby improving the reliability of results by eliminating extraneous factors associated with different animals 15,16. In the last decade, commercially bred pig varieties such as Duroc, Yorkshire, and Landrace domestic pigs 15-17 have been employed in skin wound healing studies. Despite the clinical validity and applicability of porcine models, larger breeding spaces and
Advances in Experimental Medicine and Biology, 2018
In order to develop and commercialize for the regenerative medicinal products, smart biomaterials... more In order to develop and commercialize for the regenerative medicinal products, smart biomaterials with biocompatibility must be needed. In this chapter, we introduce collagen and hyaluronic acid (HA) as extracellular matrix as well as deal with the molecular mechanism as microenvironment, mechanistic effects, and gene expression. Application of collagen and HA have been reviewed in the area of orthopedics, orthopedics, ophthalmology, dermatology and plastic surgery. Finally, the ongoing and commercial products of collagen and HA for regenerative medicine have been introduced.
Acanthamoeba is free-living protist pathogen capable of causing a blinding keratitis and granulom... more Acanthamoeba is free-living protist pathogen capable of causing a blinding keratitis and granulomatous encephalitis. However, the mechanisms of Acanthamoeba pathogenesis are still not clear. Here, our results show that cells co-cultured with pathogenic Acanthamoeba would be spherical and floated, even without contacting the protists. Then, the Acanthamoeba protists would contact and engulf these cells. In order to clarify the contact-independent pathogenesis mechanism in Acanthamoeba, we collected the Acanthamoeba-secreted proteins (Asp) to incubate with cells for identifying the extracellular virulent factors and investigating the cytotoxicity process. The Asps of pathogenic Acanthamoeba express protease activity to reactive Leu amino acid in ECM and induce cell-losing adhesion ability. The M20/M25/M40 superfamily aminopeptidase protein (ACA1_264610), an aminopeptidase be found in Asp, is upregulated after Acanthamoeba and C6 cell co-culturing for 6 h. Pre-treating the Asp with leucine aminopeptidase inhibitor and the specific antibodies of Acanthamoeba M20/M25/M40 superfamily aminopeptidase could reduce the cell damage during Asp and cell co-incubation. These results suggest an important functional role of the Acanthamoeba secreted extracellular aminopeptidases in the Acanthamoeba pathogenesis process. This study provides information regarding clinically pathogenic isolates to target specific molecules and design combined drugs.
This study has demonstrated that baicalein has anticancer effectiveness against human hepatoma ce... more This study has demonstrated that baicalein has anticancer effectiveness against human hepatoma cells. The dose response of baicalein in Hep G2 and Hep J2 cells indicates that baicalein decreased viability >90%. In comparison, baicalein had only minimal effects on the viability of control Chang liver cells. Flow cytometric analysis showed that baicalein inhibited the cell cycle of Hep G2 cells in the S phase. In addition, baicalein treatment resulted in a decreased mitochondrial transmembrane potential and damaged the integrity of the cell membrane. The TdT-mediated dUTP-biotin nick end labeling assay results indicated that baicalein elicited a significant increase of DNA fragmentation in Hep G2 cells after incubation for 48 hours. These results indicate that baicalein is an effective antihepatoma agent with minimal influence on noncancer cells. The effects of baicalein on Hep G2 cells include inhibition of the S phase of the cell cycle, dysfunction of mitochondria, and initiation of apoptosis.
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Papers by Lynn Huang