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

Abstract

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).

Tissue Engineering: Challenges and Selected Application

“…52 Despite acceptable results in epithelialization of keratinocytes with synthetic polymers, no successful epidermal graft has been achieved, due to their limited cellular recognition and tissue compatibility. Synthetic polymers in combination with natural polymers can be used for temporary dressing, epidermal/dermal cell carriers, or full-thickness skin equivalent, 53 PLGA/collagen, 54 PLA/ chitosan, 55 PCL/chitosan, 56 PCL/collagen 57 currently used for skin graft development.…”

The Influence of Tetracycline Loading on the Surface Morphology and Biocompatibility of Films Made from P(3HB) Microspheres

“…These cell lines are isolated from the periphery of a melanoma of human adult skin. [1] These differentiated keratinocytes appear to be similar to normal human keratinocytes. When the skin is subjected to injury, the keratinoyctes present near the wound site respond to the biochemical signaling by migrating and adhering to the site.…”

“…in cardiovascular surgery or replacement of damaged skin) should have adequate properties for supporting cell attachment and cell proliferation and should also possess sufficient mechanical strength. [1] In previous studies, films made of polysaccharides; a-hydroxy acids and their copolymers have been used as dermal tissue patches. However, due to some disadvantages such as prolonged healing times and limited tissue functionality, alternative biodegradable polymers have been considered.…”

“…However, due to some disadvantages such as prolonged healing times and limited tissue functionality, alternative biodegradable polymers have been considered. [1][2][3] For skin graft or wound healing studies one of the most commonly used cell types is the human adult low calcium high temperature (HaCaT) cell line. These cell lines are isolated from the periphery of a melanoma of human adult skin.…”

Highly elastomeric poly(3-hydroxyoctanoate) based natural polymer composite for enhanced keratinocyte regeneration

“…The skin cell matrix construct is then grafted on to the wound (epidermal graft); cells then proliferate from the matrix to the wound bed forming cell clusters and ultimately the normal epidermis. [14,15] The matrix thus provides healthy replacement cells, protection to the wound, until it is degraded or absorbed [14,15] , and elasticity and mechanical support to the regenerating skin. Amongst the natural materials, one promising candidate for designing the matrix support, are the polyhydroxyalkanoate (PHA) family of biopolymers [16] PHAs are polyesters of 3-hydroxyacids biosynthesized from bacteria using a variety of carbon sources.…”