Growth of keratinocytes on porous films of poly(3-hydroxybutyrate) and poly(4-hydroxybutyrate) blended with hyaluronic acid and chitosan.

The objective of this study was to develop novel absorbable films suitable for use as a tissue-engineering scaffold for keratinocytes as a therapy for replacement of damaged skin. Poly(4-hydroxybutyrate) (P(4HB)) and poly (3-hydroxybutyrate) (P(3HB)) were blended with small amounts of the polysaccharides hyaluronic acid (HA), chitosan (CH), pectin and alginic acid, and were solution cast to produce porous films. The resulting composites had favorable mechanical properties, and these films were compared with two commercially available implantable films made of poly(L-lactide-co-D,L-lactide) (PLA copolymer) and HA benzyl ester. Tensile testing demonstrated that a high level of flexibility of P(4HB) was retained in the P(4HB)-polysaccharide composite films, whereas the P(3HB) film and its polysaccharide composites were stiffer and more brittle. The proliferation kinetics of adherent HaCaT keratinocytes on the films was examined in vitro. The porous surface of the P(4HB) and P(3HB) films blended with HA or CH promoted the growth of keratinocytes significantly. The order of maximum cell numbers on these films was P(4HB)/HA > P(4HB)/CH > P(3HB)/HA > P(3HB)/CH > P(3HB)/pectin > P(3HB)/alginic acid. Scanning electron microscopy and confocal laser scanning microscopy revealed differences in cell growth. Cells formed clusters on P(3HB) and its composites, while the cells grew as a confluent layer on P(4HB) and its composites. HaCaT cells formed large numbers of filaments only on P(4HB) films, indicating the excellent biocompatibility of this material. For the nonporous PHB films, the proliferation rate of cells was found to increase with decreasing hydrophobicity in the order: P(4HB) > P(3HB)/P(4HB) blend > P(3HB).

[1]  Guoqiang Chen,et al.  The effect of D,L-beta-hydroxybutyric acid on cell death and proliferation in L929 cells. , 2006, Biomaterials.

[2]  Qiong Wu,et al.  Effect of 3-hydroxyhexanoate content in poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) on in vitro growth and differentiation of smooth muscle cells. , 2006, Biomaterials.

[3]  R. Androsch,et al.  In vitro and in vivo studies on blends of isotactic and atactic poly (3-hydroxybutyrate) for development of a dura substitute material. , 2006, Biomaterials.

[4]  Chao Yang,et al.  In vitro fabrication of a tissue engineered human cardiovascular patch for future use in cardiovascular surgery. , 2006, The Annals of thoracic surgery.

[5]  Y. Gong,et al.  Effects of surface modification of poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) on physicochemical properties and on interactions with MC3T3-E1 cells. , 2005, Journal of biomedical materials research. Part A.

[6]  T. Lim,et al.  Assimilating cell sheets and hybrid scaffolds for dermal tissue engineering. , 2005, Journal of biomedical materials research. Part A.

[7]  Guoqiang Chen,et al.  The application of polyhydroxyalkanoates as tissue engineering materials. , 2005, Biomaterials.

[8]  Guo-Qiang Chen,et al.  Effects of crystallization of polyhydroxyalkanoate blend on surface physicochemical properties and interactions with rabbit articular cartilage chondrocytes. , 2005, Biomaterials.

[9]  S. Dommerich,et al.  In-vitro-Kultivierung von Zellen der oralen Schleimhaut auf Matrizes aus Kollagen, Poly-L-Lactid (PLLA) und Polyhydroxybuttersäure (PHB) , 2005 .

[10]  A. Coombes,et al.  A co-cultured skin model based on cell support membranes. , 2005, Biochemical and biophysical research communications.

[11]  Guoqiang Chen,et al.  Influence of dl-β-Hydroxybutyric Acid on Cell Proliferation and Calcium Influx† , 2005 .

[12]  Qiong Wu,et al.  Effect of composition of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) on growth of fibroblast and osteoblast. , 2005, Biomaterials.

[13]  Ashok K. Srivastava,et al.  Recent advances in microbial polyhydroxyalkanoates , 2005 .

[14]  L. Gibson,et al.  The effect of pore size on cell adhesion in collagen-GAG scaffolds. , 2005, Biomaterials.

[15]  Y. Gong,et al.  Novel biodegradable films and scaffolds of chitosan blended with poly(3-hydroxybutyrate) , 2005, Journal of biomaterials science. Polymer edition.

[16]  D. Hutmacher,et al.  In vitro characterization of natural and synthetic dermal matrices cultured with human dermal fibroblasts. , 2004, Biomaterials.

[17]  T G Volova,et al.  Tissue response to the implantation of biodegradable polyhydroxyalkanoate sutures , 2004, Journal of materials science. Materials in medicine.

[18]  Kangde Yao,et al.  A study on a chitosan-gelatin-hyaluronic acid scaffold as artificial skin in vitro and its tissue engineering applications , 2004, Journal of biomaterials science. Polymer edition.

[19]  J. Ostwald,et al.  [In vitro culture of cells from respiratory mucosa on foils of collagen, poly-L-lactide (PLLA) and poly-3-hydroxy-butyrate (PHB)]. , 2003, Laryngo- rhino- otologie.

[20]  Chi-Hsiung Jou,et al.  Protein adsorption, fibroblast activity and antibacterial properties of poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) grafted with chitosan and chitooligosaccharide after immobilized with hyaluronic acid. , 2003, Biomaterials.

[21]  C. Zavaglia,et al.  Synthesis and characterization of poly(L-lactic acid) membranes: Studies in vivo and in vitro , 2003, Journal of materials science. Materials in medicine.

[22]  T. Chakrabarti,et al.  Preparation and characterization of biodegradable poly-3-hydroxybutyrate-starch blend films. , 2003, Bioresource technology.

[23]  Yilin Cao,et al.  Fabrication and surface modification of macroporous poly(L-lactic acid) and poly(L-lactic-co-glycolic acid) (70/30) cell scaffolds for human skin fibroblast cell culture. , 2002, Journal of biomedical materials research.

[24]  P. Hu,et al.  Study on the three-dimensional proliferation of rabbit articular cartilage-derived chondrocytes on polyhydroxyalkanoate scaffolds. , 2002, Biomaterials.

[25]  Guo-Qiang Chen,et al.  Effect of surface treatment on the biocompatibility of microbial polyhydroxyalkanoates. , 2002, Biomaterials.

[26]  B. Nebe,et al.  Structural alterations of adhesion mediating components in cells cultured on poly-beta-hydroxy butyric acid. , 2001, Biomaterials.

[27]  William H. Eaglstein,et al.  Tissue-Engineered Skin , 2001 .

[28]  David P. Martin,et al.  Fabrication of a trileaflet heart valve scaffold from a polyhydroxyalkanoate biopolyester for use in tissue engineering. , 2000, Tissue engineering.

[29]  M Fimiani,et al.  A new model for studying differentiation and growth of epidermal cultures on hyaluronan-based carrier. , 1999, Biomaterials.

[30]  R. Guidoin,et al.  In vivo biocompatibility and degradation studies of polyhydroxyoctanoate in the rat: a new sealant for the polyester arterial prosthesis. , 1999, Tissue engineering.

[31]  David P. Martin,et al.  PHA applications: addressing the price performance issue: I. Tissue engineering. , 1999, International journal of biological macromolecules.

[32]  V. Terskikh,et al.  Cultivation and transplantation of epidermal keratinocytes. , 1999, International review of cytology.

[33]  Río,et al.  Axisymmetric Drop Shape Analysis: Computational Methods for the Measurement of Interfacial Properties from the Shape and Dimensions of Pendant and Sessile Drops. , 1997, Journal of colloid and interface science.

[34]  J. Hornung,et al.  Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line , 1988, The Journal of cell biology.