By intercepting the most effective fragment of human collagen’s gene,recombinant human-like collagen is produced by genetic engineering technology and biotechnology.The collagen has good biocompatibility and can prom...By intercepting the most effective fragment of human collagen’s gene,recombinant human-like collagen is produced by genetic engineering technology and biotechnology.The collagen has good biocompatibility and can promote cell formation.Recombinant human-like collagen has great potential application value in medical biological materials,cosmetics and foods.In recent years,many specialized enterprises which are engaged in human-like collagen raw materials production have emerged.In this paper,a brief summary of the current recombinant human-like collagen raw material’s production and its latest research and development have been made.展开更多
We reported recombinant human-like type I collagen inducing growth of hydroxyapatite crystals in vitro in the form of self-assembly of nano-fibrils of mineralized collagen resembling extracellular matrix, which obey t...We reported recombinant human-like type I collagen inducing growth of hydroxyapatite crystals in vitro in the form of self-assembly of nano-fibrils of mineralized collagen resembling extracellular matrix, which obey the same rules, but is superior to the collagen derived from animal tissues because the latter may carry diseases of animals and cause immunological reactions. The mineralized collagen fibrils aligned parallel to each other to form mineralized collagen fibers. Hydroxyapatite nanocrystals grew on the surface of these collagen fibrils with the c-axis of nanocrystals of HA orienting along the longitudinal axis of the fibrils.展开更多
Studying the skin care efficacy of recombinant humanized collagen based on in vitro level.The stability of the recombinant humanized collagen was first analyzed by treating at different temperatures,then its skincare ...Studying the skin care efficacy of recombinant humanized collagen based on in vitro level.The stability of the recombinant humanized collagen was first analyzed by treating at different temperatures,then its skincare efficacy based on in vitro level was evaluated by detecting the inhibition rate of elastase,the inhibition rate of collagenase,the protein content of type I collagen in human fibroblasts,the inhibition of reactive oxygen species(ROS)with human keratinocytes,and the effects of the recombinant humanized collagen on the expression of hyaluronic acid(HA),filaggrin(FLG)and transglutaminase 1(TGM1)in keratinocytes.The results showed that recombinant humanized collagen was able to maintain stability at temperatures below 70℃.With regard to its skincare efficacy,recombinant humanized collagen could inhibit elastase and collagenase activities and promote the increase of type I collagen content in human fibroblasts.It also showed good inhibition of ROS in keratinocytes in vitro and could increase the expression of HA,FLG,and TGM1 in keratinocytes.In short,the recombinant humanized collagen exhibited a favourable skin care effect in vitro level.This study proved that it has potential firming,anti-wrinkle,moisturizing,and repairing efficacy,and is a valuable cosmetic raw material.展开更多
In order to improve the production of human-like collagen III(HLC III)by fed-batch culture of recombinant Escherichia coli BL21,the Plackett-Burman and Box-Behnken design were applied to optimize the fermentation proc...In order to improve the production of human-like collagen III(HLC III)by fed-batch culture of recombinant Escherichia coli BL21,the Plackett-Burman and Box-Behnken design were applied to optimize the fermentation process parameters.Three variables(induction time,inoculum age and pH),which have significant effects on HLC III production,were selected from eight variables by Plackett-Burman design.With the regression coefficient analysis in the Box-Behnken design,a relationship between HLC III production and three significant factors was obtained,and the optimum levels of the three variables were as follows:induction time 3.2h,inoculum age 12.6 h and pH 6.7.The 3D response surface plots and 2D contour plots created by the Box-Behnken design showed that the interaction between induction time and pH and that between innoculum age and pH were significant.An average 9.68 g·L1HLC III production was attained in the validation experiment under optimized condition,which was 80%higher than the yield of 5.36 g·L1before optimization.展开更多
The kinetics of batch and fed-batch cultures of recombinant Escherichia coli producing human-like collagen was investigated. In the batch culture, a kinetic model of a simple growth-association system was concluded wi...The kinetics of batch and fed-batch cultures of recombinant Escherichia coli producing human-like collagen was investigated. In the batch culture, a kinetic model of a simple growth-association system was concluded without consideration of cell endogeneous metabolism. The cell lag time, the maximum specific growth rate and Yx/s were determined as 1.75h, 0.65h^-1 and 0.51g·g^-1, respectively. In the fed-batch culture, different specific growth rates were set at （0.15, 0.2, 0.25h^-1） by the method of pseudo-exponential feeding, and the expressions for the specific rate of substrate consumption, the growth kinetics and the product formation kinetics of each phase were obtained. The result shows that the concentrations of cell and product can reach 77.5g·L^-1 and 10.2g·L^-1 respectively. The modal predictions are in good agreement with the experimental data.展开更多
Recombinant Escherichia coli BL21 is used to produce human-like collagen. The key constituents of media are optimized using response surface methodology (RSM). Before thermal induction, the highest biomass production ...Recombinant Escherichia coli BL21 is used to produce human-like collagen. The key constituents of media are optimized using response surface methodology (RSM). Before thermal induction, the highest biomass production and the lowest production of some hazardous by-products, especially acetic acid, were obtained in the media containing 0.085 mol·L-1 glucose and 0.019 mol·L-1 nitrogen (carbon-nitrogen ratio, 4.47:1). After thermal induction, when the concentrations of glucose and nitrogen in the media were 0.065 mol·L-1 and 0.017 mol·L-1 , respectively (carbon-nitrogen ratio, 3.82:1), the productivity of human-like collagen per cell was the highest while that of acetic acid was the lowest. The extended analysis showed that the production of lactic acid and propionic acid increased while that of some intermediate acids of the tricarboxylic acid cycle decreased if the dose of glucose increased.展开更多
Recombinant E. coli BL 21 was cultivated in high cell density to produce human-like collagen. The effects of the feeding of nitrogen source, controlled by an auto on/off-feeding mode with two different cycles of 0.5mi...Recombinant E. coli BL 21 was cultivated in high cell density to produce human-like collagen. The effects of the feeding of nitrogen source, controlled by an auto on/off-feeding mode with two different cycles of 0.5min and 4min intervals, oxygen-enrichment methods and inducement strength on the cell yield and human-like collagen production were investigated. The studies showed that nitrogen source feeding in fast cycle could result in higher human-like collagen production than that in slow cycle; and the feedback regulation of glucose, increase of the pressure of fermentation bioreactor, and supply of oxygen-enriched air could all increase cell yield and human-like collagen production. The effects of inducement strength on protein expression were found important. When OD600 reached 90-100, the cultivation temperature was increased to 42℃ to begin induction for 2-3 h, and then shifted to 39℃ for 5-6h induction, the cell density and human-like collagen production could reach 96g·L-1 [DCW (dry cell mass)] and 19.8% (g·g-1 DCW) respectively.展开更多
Recombinant collagen is a pivotal topic in foundational biological research and epitomizes the application of critical bioengineer-ing technologies.These technological advancements have pro-found implications across d...Recombinant collagen is a pivotal topic in foundational biological research and epitomizes the application of critical bioengineer-ing technologies.These technological advancements have pro-found implications across diverse areas such as regenerative medicine,organ replacement,tissue engineering,cosmetics and more.Thus,recombinant collagen and its preparation methodologies rooted in genetically engineered celis mark pivotal milestones in medical product research.This article pro-vides a comprehensive overview of the current genetic engi-neering technologies and methods used in the production of recombinant collagen,as well as the conventional production process and gquality control detection methods for this material.Furthermore,the discussion extends to foresee the strides in physical transfection and magnetic control sorting studies,envisioning an enhanced preparation of recombinant collagen-seeded cells to further fuel recombinant collagen production.展开更多
The management of chronic wounds in diabetes remains challenging due to the complexity of impaired wound healing,delayed healing,susceptibility to infection,and elevated risk of reopening,highlighting the need for eff...The management of chronic wounds in diabetes remains challenging due to the complexity of impaired wound healing,delayed healing,susceptibility to infection,and elevated risk of reopening,highlighting the need for effective chronic wound management with innovative approaches such as multifunctional hydrogels.Here,we have produced HA-DA@rhCol hydrogels consisting of dopamine-modified hyaluronic acid and recombinant human collagen type-III(rhCol)by oxidative coupling of the catechol group using the H_(2)O_(2)/HRP catalytic system.The post-reactive hydrogel has a good porous structure,swelling rate,reasonable degradation,rheological and mechanical properties,and the catechol group and dopamine impart to the hydrogel tissue adhesiveness,antioxidant capacity,and excellent photothermal effects leading to superior in vitro antimicrobial activity.In addition,the ability of rhCol to confer hydrogels to promote angiogenesis and wound repair has also been investigated.Cytotoxicity and hemolysis tests demonstrated the good biocompatibility of the hydrogel.Wound closure,collagen deposition and immunohistochemical examination confirmed the ability of the hydrogel to promote diabetic wound healing.In summary,the adhesive hemostatic antioxidative hydrogel with rhCol to promote wound healing in diabetic rat is an excellent chronic wound dressing.展开更多
Collagens are the most abundant proteins in mammals and form an extracellular matrix (ECM) with other components as the structural support of muscle, skin, corneas and blood vessels etc. Other than providing structura...Collagens are the most abundant proteins in mammals and form an extracellular matrix (ECM) with other components as the structural support of muscle, skin, corneas and blood vessels etc. Other than providing structural support, the ECM exhibits active communication with cells and influences many cellular processes including migration, wound healing, differentiation and cancer metastasis. Though collagen proteins contain highly repetitive primary sequences and defined tertiary structures, more and more studies have shown that many short peptides/motifs within collagen proteins play key roles in various biological processes. These short sequences are effective within triple helical structures or independently as stand-alone molecules resulting from proteolytic degradation. Besides endogenous ECM-derived peptides, many more functional peptides have been produced by tissue processing, chemical synthesis, and recombinant protein production. In this review, we summarize different peptides/motifs identified in collagen and other ECM proteins and discuss their potential for medical, personal care, and cosmetics applications.展开更多
Recently,evidence has suggested that chronic endometritis(CE)is a crucial factor associated with infertility and failure of assisted reproductive techniques,prompting concern in the reproductive field.Studies have sho...Recently,evidence has suggested that chronic endometritis(CE)is a crucial factor associated with infertility and failure of assisted reproductive techniques,prompting concern in the reproductive field.Studies have shown that persistent infiltered immune cells stimulation result in the disturbance of endometrial immune microenvironment could lead to the infertility of CE patients finally.Conventional treatments are limited because they lack immune regulation,so it is urgent to develop a novel approach to treat CE and promote embryo implantation in patients with CE.Herein,we prepared recombinant humanized type III collagen(rhCol III)with high cell adhesion activity to regulate macrophages and repair the endometrium.In this study,M1 macrophages and M1 macrophages cultured medium and lipopolysaccharide(LPS)co-stimulated inflammatory endometrium stromal cells(ESCs)were established in vitro to mimic CE condition.rhCol III promoted M1 macrophages toward M2 phenotype,improved cell migration,viability and collagen components of inflammatory ESCs.Also,the inflammatory response of inflammatory ESCs was downregulated after rhCol III treatment.Subsequently,LPS was used for CE rat model and a 28-day observation was performed;inflammatory cells’infiltration,endometrium repair,extracellular matrix(ECM)remodeling and pregnancy outcomes were promoted after rhCol III endometrial infusion.In conclusion,rhCol III promoted(i)macrophage polarization toward M2 macrophages,(ii)pro-inflammatory cytokine production and anti-inflammatory cytokine reduction,(iii)ECM remodeling and(iv)fertility restoration.Meanwhile,rhCol III enhanced cell biological functions by interacting with discoidin domain receptors,regulated cell metabolism and reduced the inflammatory response through the inhibition of the NF-κB/YAP signaling pathway.Overall,the results illustrated the potential therapeutic prospects of rhCol III for CE treatment.展开更多
文摘By intercepting the most effective fragment of human collagen’s gene,recombinant human-like collagen is produced by genetic engineering technology and biotechnology.The collagen has good biocompatibility and can promote cell formation.Recombinant human-like collagen has great potential application value in medical biological materials,cosmetics and foods.In recent years,many specialized enterprises which are engaged in human-like collagen raw materials production have emerged.In this paper,a brief summary of the current recombinant human-like collagen raw material’s production and its latest research and development have been made.
文摘We reported recombinant human-like type I collagen inducing growth of hydroxyapatite crystals in vitro in the form of self-assembly of nano-fibrils of mineralized collagen resembling extracellular matrix, which obey the same rules, but is superior to the collagen derived from animal tissues because the latter may carry diseases of animals and cause immunological reactions. The mineralized collagen fibrils aligned parallel to each other to form mineralized collagen fibers. Hydroxyapatite nanocrystals grew on the surface of these collagen fibrils with the c-axis of nanocrystals of HA orienting along the longitudinal axis of the fibrils.
文摘Studying the skin care efficacy of recombinant humanized collagen based on in vitro level.The stability of the recombinant humanized collagen was first analyzed by treating at different temperatures,then its skincare efficacy based on in vitro level was evaluated by detecting the inhibition rate of elastase,the inhibition rate of collagenase,the protein content of type I collagen in human fibroblasts,the inhibition of reactive oxygen species(ROS)with human keratinocytes,and the effects of the recombinant humanized collagen on the expression of hyaluronic acid(HA),filaggrin(FLG)and transglutaminase 1(TGM1)in keratinocytes.The results showed that recombinant humanized collagen was able to maintain stability at temperatures below 70℃.With regard to its skincare efficacy,recombinant humanized collagen could inhibit elastase and collagenase activities and promote the increase of type I collagen content in human fibroblasts.It also showed good inhibition of ROS in keratinocytes in vitro and could increase the expression of HA,FLG,and TGM1 in keratinocytes.In short,the recombinant humanized collagen exhibited a favourable skin care effect in vitro level.This study proved that it has potential firming,anti-wrinkle,moisturizing,and repairing efficacy,and is a valuable cosmetic raw material.
基金Supported by the National Natural Science Foundation of China（20776119） the National High Technology Research and Development Program of China（2007AA03Z456A） the Special Research Program of the Education Department of Shaanxi Province（07JK417）
文摘In order to improve the production of human-like collagen III(HLC III)by fed-batch culture of recombinant Escherichia coli BL21,the Plackett-Burman and Box-Behnken design were applied to optimize the fermentation process parameters.Three variables(induction time,inoculum age and pH),which have significant effects on HLC III production,were selected from eight variables by Plackett-Burman design.With the regression coefficient analysis in the Box-Behnken design,a relationship between HLC III production and three significant factors was obtained,and the optimum levels of the three variables were as follows:induction time 3.2h,inoculum age 12.6 h and pH 6.7.The 3D response surface plots and 2D contour plots created by the Box-Behnken design showed that the interaction between induction time and pH and that between innoculum age and pH were significant.An average 9.68 g·L1HLC III production was attained in the validation experiment under optimized condition,which was 80%higher than the yield of 5.36 g·L1before optimization.
基金Supported by the National Science and Technology Key Funds (2003DA901A32) and the National Natural Science Foundationof China (No.20476085).
文摘The kinetics of batch and fed-batch cultures of recombinant Escherichia coli producing human-like collagen was investigated. In the batch culture, a kinetic model of a simple growth-association system was concluded without consideration of cell endogeneous metabolism. The cell lag time, the maximum specific growth rate and Yx/s were determined as 1.75h, 0.65h^-1 and 0.51g·g^-1, respectively. In the fed-batch culture, different specific growth rates were set at （0.15, 0.2, 0.25h^-1） by the method of pseudo-exponential feeding, and the expressions for the specific rate of substrate consumption, the growth kinetics and the product formation kinetics of each phase were obtained. The result shows that the concentrations of cell and product can reach 77.5g·L^-1 and 10.2g·L^-1 respectively. The modal predictions are in good agreement with the experimental data.
基金Supported by the National High Technology Research and Development Program of China （2006AA02Z246 2007AA03Z456） the National Natural Science Foundation of China （20776119 21076169）+4 种基金 Xi’an Research and Development Program（CX0735） the Scientific Research Program of Shaanxi Provincial Department of Education China （07JK417 07JC16 JG08181） the Natural Science Foundation of Shaanxi Province （2010JQ2012） the Specialized Research Fund for the Doctoral Program of Higher Education of China （20096101120023 20096101110014） Shaanxi Key Subject Program China
文摘Recombinant Escherichia coli BL21 is used to produce human-like collagen. The key constituents of media are optimized using response surface methodology (RSM). Before thermal induction, the highest biomass production and the lowest production of some hazardous by-products, especially acetic acid, were obtained in the media containing 0.085 mol·L-1 glucose and 0.019 mol·L-1 nitrogen (carbon-nitrogen ratio, 4.47:1). After thermal induction, when the concentrations of glucose and nitrogen in the media were 0.065 mol·L-1 and 0.017 mol·L-1 , respectively (carbon-nitrogen ratio, 3.82:1), the productivity of human-like collagen per cell was the highest while that of acetic acid was the lowest. The extended analysis showed that the production of lactic acid and propionic acid increased while that of some intermediate acids of the tricarboxylic acid cycle decreased if the dose of glucose increased.
基金Supported by the National Science and Technology Key Funds (2003DA901A32)the National Nature Science Foundation (No. 20476085).
文摘Recombinant E. coli BL 21 was cultivated in high cell density to produce human-like collagen. The effects of the feeding of nitrogen source, controlled by an auto on/off-feeding mode with two different cycles of 0.5min and 4min intervals, oxygen-enrichment methods and inducement strength on the cell yield and human-like collagen production were investigated. The studies showed that nitrogen source feeding in fast cycle could result in higher human-like collagen production than that in slow cycle; and the feedback regulation of glucose, increase of the pressure of fermentation bioreactor, and supply of oxygen-enriched air could all increase cell yield and human-like collagen production. The effects of inducement strength on protein expression were found important. When OD600 reached 90-100, the cultivation temperature was increased to 42℃ to begin induction for 2-3 h, and then shifted to 39℃ for 5-6h induction, the cell density and human-like collagen production could reach 96g·L-1 [DCW (dry cell mass)] and 19.8% (g·g-1 DCW) respectively.
基金supported by the second batch of the China Drug Regulatory Science Action Plan(Research on safety and effectiveness evaluation of novel biomaterials)from National Medical Products Administration(NMPA).
文摘Recombinant collagen is a pivotal topic in foundational biological research and epitomizes the application of critical bioengineer-ing technologies.These technological advancements have pro-found implications across diverse areas such as regenerative medicine,organ replacement,tissue engineering,cosmetics and more.Thus,recombinant collagen and its preparation methodologies rooted in genetically engineered celis mark pivotal milestones in medical product research.This article pro-vides a comprehensive overview of the current genetic engi-neering technologies and methods used in the production of recombinant collagen,as well as the conventional production process and gquality control detection methods for this material.Furthermore,the discussion extends to foresee the strides in physical transfection and magnetic control sorting studies,envisioning an enhanced preparation of recombinant collagen-seeded cells to further fuel recombinant collagen production.
基金supported by grants from National Natural Science Foundation of China No.82172206Zhejiang Provincial Medical and Healthy Science Foundation of China(No.2023RC183 and 2024KY110).
文摘The management of chronic wounds in diabetes remains challenging due to the complexity of impaired wound healing,delayed healing,susceptibility to infection,and elevated risk of reopening,highlighting the need for effective chronic wound management with innovative approaches such as multifunctional hydrogels.Here,we have produced HA-DA@rhCol hydrogels consisting of dopamine-modified hyaluronic acid and recombinant human collagen type-III(rhCol)by oxidative coupling of the catechol group using the H_(2)O_(2)/HRP catalytic system.The post-reactive hydrogel has a good porous structure,swelling rate,reasonable degradation,rheological and mechanical properties,and the catechol group and dopamine impart to the hydrogel tissue adhesiveness,antioxidant capacity,and excellent photothermal effects leading to superior in vitro antimicrobial activity.In addition,the ability of rhCol to confer hydrogels to promote angiogenesis and wound repair has also been investigated.Cytotoxicity and hemolysis tests demonstrated the good biocompatibility of the hydrogel.Wound closure,collagen deposition and immunohistochemical examination confirmed the ability of the hydrogel to promote diabetic wound healing.In summary,the adhesive hemostatic antioxidative hydrogel with rhCol to promote wound healing in diabetic rat is an excellent chronic wound dressing.
文摘Collagens are the most abundant proteins in mammals and form an extracellular matrix (ECM) with other components as the structural support of muscle, skin, corneas and blood vessels etc. Other than providing structural support, the ECM exhibits active communication with cells and influences many cellular processes including migration, wound healing, differentiation and cancer metastasis. Though collagen proteins contain highly repetitive primary sequences and defined tertiary structures, more and more studies have shown that many short peptides/motifs within collagen proteins play key roles in various biological processes. These short sequences are effective within triple helical structures or independently as stand-alone molecules resulting from proteolytic degradation. Besides endogenous ECM-derived peptides, many more functional peptides have been produced by tissue processing, chemical synthesis, and recombinant protein production. In this review, we summarize different peptides/motifs identified in collagen and other ECM proteins and discuss their potential for medical, personal care, and cosmetics applications.
基金supported by Scientific and Technological Research Program of Chongqing Municipal Education Commission(Grant No.KJCXZD2020017).
文摘Recently,evidence has suggested that chronic endometritis(CE)is a crucial factor associated with infertility and failure of assisted reproductive techniques,prompting concern in the reproductive field.Studies have shown that persistent infiltered immune cells stimulation result in the disturbance of endometrial immune microenvironment could lead to the infertility of CE patients finally.Conventional treatments are limited because they lack immune regulation,so it is urgent to develop a novel approach to treat CE and promote embryo implantation in patients with CE.Herein,we prepared recombinant humanized type III collagen(rhCol III)with high cell adhesion activity to regulate macrophages and repair the endometrium.In this study,M1 macrophages and M1 macrophages cultured medium and lipopolysaccharide(LPS)co-stimulated inflammatory endometrium stromal cells(ESCs)were established in vitro to mimic CE condition.rhCol III promoted M1 macrophages toward M2 phenotype,improved cell migration,viability and collagen components of inflammatory ESCs.Also,the inflammatory response of inflammatory ESCs was downregulated after rhCol III treatment.Subsequently,LPS was used for CE rat model and a 28-day observation was performed;inflammatory cells’infiltration,endometrium repair,extracellular matrix(ECM)remodeling and pregnancy outcomes were promoted after rhCol III endometrial infusion.In conclusion,rhCol III promoted(i)macrophage polarization toward M2 macrophages,(ii)pro-inflammatory cytokine production and anti-inflammatory cytokine reduction,(iii)ECM remodeling and(iv)fertility restoration.Meanwhile,rhCol III enhanced cell biological functions by interacting with discoidin domain receptors,regulated cell metabolism and reduced the inflammatory response through the inhibition of the NF-κB/YAP signaling pathway.Overall,the results illustrated the potential therapeutic prospects of rhCol III for CE treatment.
