Preadipocyte Seeded PLGA Scaffolds for Adipose Tissue Engineering

Abstract

Adipose tissue equivalents have not been addressed as yet despite the clinical need in congenital deformities, posttraumatic repair, cancer rehabilitation, and other soft tissue defects. Preadipocytes were successfully harvested from rat epididymal fat pads of Sprague-Dawley and Lewis rats and expanded ex vivo. In vitro cultures demonstrated full differentiation of preadipocytes into mature adipocytes with normal lipogenic activity. The onset of differentiation was well-controlled by regulating preadipocyte confluency. Poly(lactic-co-glycolic) acid (PLGA) polymer disks with 90% porosity, 2.5 mm thick, 12 mm diameter, pore size range of 135-633 microm were fabricated and seeded with preadipocytes at 10(5) cells/mL. Disks in vitro demonstrated fully differentiated mature adipocytes within the pores of the disks. Short-term in vivo experiments were conducted by implanting preseeded disks subcutaneously on the flanks of rats for 2 and 5 weeks. Histologic staining of harvested disks with osmium tetroxide (OsO4) revealed the formation of adipose tissue throughout the disks. Fluorescence labeling of preadipocytes confirmed that formed adipose tissue originated from seeded preadipocytes rather than from possible infiltrating perivascular tissue. This study demonstrates the potential of using primary preadipocytes as a cell source in cell-seeded polymer scaffolds for tissue engineering applications.

Human clinical experience with adipose precursor cells seeded on hyaluronic acid-based spongy scaffolds

“…The three-dimensional geometry of the native ECM constrains cells during regeneration and provides space for tissue development [25] and in vitro studies have shown that the scaffold geometry promotes preadipocyte differentiation [31]. The architecture of adipose tissue has been reconstructed using adipose-derived stromal cells seeded onto different scaffold materials [6][7][8]13,14]. Hyaluronic acid-based scaffolds have been used clinically to successfully direct chondrocyte development [32][33][34].…”

“…Materials that meet fundamental requirements such as controlled degradation, cytotoxicity and immunogenity are needed. Research reports have shown successful and reproducible inoculation and culturing of adipocyteand preadipocyte-precursor cells (preadipocytes and progenitors, respectively) on synthetic [6,7] or natural [8] prefabricated scaffolds [9] with subsequent differentiation in vitro or adipose tissue formation in vivo [10]. The scaffolds within these bio-hybrids provided the seeded cells specific attachment or binding sites, shaped the tissue construct and functioned as geometrical environments in which not only cells received their essential cues for a structural cell organization and cellular behaviour but also cellular function was modulated [11,12].…”

“…The scaffolds within these bio-hybrids provided the seeded cells specific attachment or binding sites, shaped the tissue construct and functioned as geometrical environments in which not only cells received their essential cues for a structural cell organization and cellular behaviour but also cellular function was modulated [11,12]. Various scaffold materials have been analyzed in experiments for autotransplantation as well as for in vitro or in vivo growth and differentiation of preadipocytes [6,9,13,14] but no data extrapolations have been realized yet to clinical applications. In this pilot clinical trial hyaluronic acid-based (HA) scaffolds (HYAFF Ò 11) were used as a cell-carrier material.…”

Evolution and Trends in Reconstructive Facial Surgery: An Update

“…The patient will invariably experience additional trauma, pain, scarring, disfigurement or other donor site complications [50][51][52]. In addition, it has been noted that though autologous grafts and flaps do a good job of replacing skeletal and soft tissue defects, they may not adequately restore the delicate complex facial anatomy [2].…”

“…The selection criteria for both recipient and donor must include physical characteristics (skin color, gender), immunological characteristics (immune status, blood type, HLA, previous sensitization), psychological characteristics (willingness to undergo the procedure, and to accept lifelong immunosuppression, coping skills, expectations), social characteristics (family support, religious beliefs, etc.) and economic status (ability to sustain prolong medical care) [2,50,51].…”