Chong Alphonsus K S, Riboh Jonathan, Smith R Lane, Lindsey Derek P, Pham Hung M, Chang James
Stanford and Palo Alto, Calif. From the Division of Plastic and Reconstructive Surgery, Stanford University Medical Center; the Bone and Joint Rehabilitation Research and Development Center, Veterans Affairs Palo Alto Health Care System; and the Department of Functional Restoration, Stanford School of Medicine.
Plast Reconstr Surg. 2009 Jun;123(6):1759-1766. doi: 10.1097/PRS.0b013e3181a65ae7.
Tissue engineering of flexor tendons requires scaffolds with adequate strength and biocompatibility. The biomechanical properties of acellularized and reseeded flexor tendon scaffolds are unknown. Acellularized tendons and reseeded constructs were tested to determine whether the treatment process had altered their biomechanical properties.
Rabbit flexor tendons were acellularized using a freeze-thaw cycle followed by trypsin and Triton-X treatment. Complete acellularization of the tendon samples was confirmed by histology and by attempting to obtain viable cells by trypsin treatment of acellularized tendon. Reseeded constructs were obtained by incubating acellularized tendons in a tenocyte suspension. Tensile testing was performed to compare the ultimate tensile stress and elastic modulus of acellularized tendons and reseeded flexor tendon constructs to control flexor tendons.
The treatment protocol successfully acellularized flexor tendons. No cells were seen within the tendon on histologic assessment, and no viable cells could be obtained from acellularized tendon. Acellularized tendon was successfully reseeded with tenocytes, although cell adhesion was limited to the surface of the tendon scaffold. Tensile testing showed that acellularized tendon had the same ultimate stress and elastic modulus as normal tendons. Reseeded tendons had the same elastic modulus as normal tendons, but hind-paw tendon constructs showed a decrease in ultimate stress compared with normal tendons (50.09 MPa versus 66.01 MPa, p = 0.026).
Acellularized flexor tendons are a potential high-strength scaffold for flexor tendon tissue engineering. This approach of acellularization and reseeding of flexor tendons may provide additional intrasynovial graft material for hand reconstruction.
屈肌腱组织工程需要具有足够强度和生物相容性的支架。脱细胞并重新接种的屈肌腱支架的生物力学特性尚不清楚。对脱细胞肌腱和重新接种的构建体进行测试,以确定处理过程是否改变了它们的生物力学特性。
采用冻融循环,随后用胰蛋白酶和 Triton-X 处理使兔屈肌腱脱细胞。通过组织学检查以及尝试通过对脱细胞肌腱进行胰蛋白酶处理来获得活细胞,确认肌腱样本完全脱细胞。通过将脱细胞肌腱在肌腱细胞悬液中孵育获得重新接种的构建体。进行拉伸试验,以比较脱细胞肌腱和重新接种的屈肌腱构建体与对照屈肌腱的极限拉伸应力和弹性模量。
该处理方案成功使屈肌腱脱细胞。组织学评估未在肌腱内发现细胞,且无法从脱细胞肌腱中获得活细胞。脱细胞肌腱成功地重新接种了肌腱细胞,尽管细胞黏附仅限于肌腱支架表面。拉伸试验表明,脱细胞肌腱的极限应力和弹性模量与正常肌腱相同。重新接种的肌腱与正常肌腱具有相同的弹性模量,但后爪肌腱构建体与正常肌腱相比极限应力降低(50.09 MPa 对 66.01 MPa,p = 0.026)。
脱细胞屈肌腱是屈肌腱组织工程潜在的高强度支架。这种屈肌腱脱细胞和重新接种的方法可为手部重建提供额外的滑膜内移植材料。