Peripheral nerve autografts to the rat spinal cord: Studies with axonal tracing methods

In young adult female rats, autologous sciatic nerve segments were transplanted to the thoracic region of the spinal cord. The grafts became well innervated but led to no obvious functional improvement. The origin and termination of axons in the grafts was studied by retrograde neuronal labeling with horseradish peroxidase (HRP) and radioautographic axonal tracing. Studies with HRP indicated that some axons in the grafts originated from intrinsic CNS neurons with their cell bodies in nearby segments of the spinal cord and that others arose from dorsal root ganglia at the level of the grafts and at least 7 segments distal to them. After tritiated amino acids were injected into lumbar dorsal root ganglia, labeled axons could be followed into the grafts but not into the rostral spinal cord stumps. Together with other experimental observations, these results demonstrate a correlation between success or failure of elongation of dorsal root fibers and peripheral or central ensheathment at the axonal tip. The corticospinal tract was studied both with radioautography and retrograde axonal transport of HRP but no extension of its axons into peripheral nerve grafts was detected under these experimental conditions. The findings implicate both neuroglial and axonal factors in the feeble regenerative response usually seen after injury to the spinal cord.     

Intracerebral grafting of cultured autologous skin fibroblasts into the rat striatum: an assessment of graft size and ultrastructure

To identify a suitable donor cell population for gene therapy applications to the central nervous system, primary fibroblasts isolated from skin biopsies and maintained in culture are employed as autologous cells for intracerebral grafting within the adult rat striatum. Results from the present investigation reveal that cultured primary skin fibroblasts cease to proliferate once they reach confluence; these cells are thus contact inhibited in vitro. Following implantation within the striatum, the volume of the primary fibroblast grafts, stained immunohistochemically for fibronectin, does not differ significantly at 3 and 8 weeks. The graft size is dependent on the density of the cell suspension, but not dependent on either the number of passages the cells are taken through in culture prior to grafting or on the postoperative survival period. Ultrastructural evidence reveals that at 8 weeks the grafts are composed primarily of collagen and fibroblasts with rough endoplasmic reticulum and vesicles. Reactive astrocytic processes and phagocytic cells are also present in the grafts. The grafts are extensively vascularized with capillaries composed of nonfenestrated endothelium; intercellular junctions are evident at sites of apposition between endothelial cells. It is concluded that primary skin fibroblasts are able to survive for at least 8 weeks following intracerebral implantation and continue to synthesize collagen and fibronectin in vivo. Also, the grafts maintain a constant volume between 3 and 8 weeks, thereby indicating that primary skin fibroblasts do not produce tumors. Finally, dynamic host-to-graft interactions--including phagocytic migration, astrocytic hypertrophy and infiltration within the grafts, and angiogenesis--are features that constitute the structural integration of primary skin fibroblasts grafted within the adult rat central nervous system.     

缺氧诱导因子1α与Gax基因对自体静脉移植术后内膜过度增生的调控机制

自体静脉移植术后内膜过度增生是影响术后效果的重要原因,缺氧诱导因子1α(HIF-1α)与Gax基因可以抑制自体静脉移植术后内膜过度增生,可作为治疗移植静脉再狭窄的一个新手段,HIF-1α通过调控血管内皮生长因子的表达,对恢复血管内皮细胞的结构和功能起着重要的调节作用,而Gax基因通过影响细胞周期和诱导细胞凋亡两条途径来调节血管平滑肌细胞的增殖。     

Involvement of melanocortin-1 receptor in the hyperpigmentation of human skin autografts

Hyperpigmentation frequently occurs in human skin autografts resulting in an unsatisfactory appearance. This study aimed to elucidate the role of melanocortin-1 receptor in the hyperpigmentation process of skin autografts by analyzing the expression of melanocortin-1 receptor. The data were correlated with the amount of melanin in autografted human skin and normal skin determined in a previous study. Immunohistochemistry, western blotting and quantitative real-time polymerase chain reaction were carried out to detect the expression and distribution of melanocortin-1 receptor in skin autografts including full-thickness skin autografts, split-thickness skin autografts and normal full-thickness skin. Fontana-Masson stain was used to detect melanin in all types of skin specimens. The expression level of melanocortin-1 receptor in autografted skin was much higher than that in control normal skin, and thinner split-thickness skin autografts had higher levels of expression of melanocortin-1 receptor than thicker grafts. The amount of melanin in skin autografts was significantly increased compared with normal skin. The expression of melanocortin-1 receptor correlated well with the amount of melanin in the epidermis of skin autografts. These results indicate that melanogenesis is dramatically enhanced in skin autografts by the melanocortin-1 receptor, and suggest that overexpression of melanocortin-1 receptor may play an important role in the hyperpigmented process of skin autografts. This study provides a novel mechanism for hyperpigmentation in skin autografts.     

纤维蛋白胶在翼状胬肉手术中应用的研究进展

翼状胬肉是一种最常见的眼表疾病之一,覆盖角膜进而影响视力和美观。临床治疗以胬肉切除联合自体结膜移植手术为主,术中结膜缝合既费时又容易引起眼部刺激、眼红等并发症。为了预防这些并发症,学者们尝试使用纤维蛋白胶替代缝线显示了独特优势。本文就纤维蛋白胶在翼状胬肉手术中应用的优缺点和研究进展做一综述,为临床应用提供依据。     

Intercalary reconstruction following resection of diaphyseal bone tumors: A systematic review

IntroductionThe options for the reconstruction of diaphyseal defects following the resection of bone tumors include biological or prosthetic implants. The purpose of our study was to evaluate different types of intercalary reconstruction techniques, including massive bone allograft, extracorporeal devitalized autograft, vascularized free fibula, and modular prosthesis.MethodsWe performed a systematic review of articles using the terms diaphyseal bone tumor and intercalary reconstruction. All the studies reporting the non-oncological complications such as infection, nonunion and fracture of the intercalary reconstructions were included. We excluded articles published before 2000 or did not involve humans in the study. Case reports, reviews, technique notes and opinion articles were also excluded based on the abstracts. Thirty-three articles included in this review were then studied to evaluate failure rates, complications and functional outcome of different surgical intercalary reconstruction techniques.ResultsNonunion rates of allograft ranged 6%–43%, while aseptic loosening rates of modular prosthesis ranged 0%–33%. Nonunion rates of allograft alone and allograft with a vascularized fibula graft ranged 6%–43% and 0%–33%, respectively. Fracture rates of allograft alone and allograft with a vascularized fibula graft ranged 7%–45% and 0%–44%, respectively. Infection rates of allograft alone and allograft with a vascularized fibula graft ranged 0%–28% and 0%–17%, respectively. All of the allograft (range: 67%–92%), extracorporeal devitalized autograft including irradiation (87%), autoclaving (70%), pasteurization (88%), low-heat (90%) or freezing with liquid nitrogen (90%), and modular prosthesis (range: 77%–93%) had similar Musculoskeletal Tumor Society functional scores. Addition of a vascularized fibula graft to allograft did not affect functional outcome [allograft with a vascularized fibula graft (range: 86%–94%) vs. allograft alone (range: 67%–92%)].ConclusionAseptic loosening rates of modular prosthesis seem to be less than nonunion rates of allograft. Adding a vascularized fibula graft to allograft seems to increase bone union rate and reduce the risk of fractures and infections, though a vascularized fibula graft needs longer surgical time and has the disadvantage of donor site morbidity. These various intercalary reconstruction techniques with or without a vascularized fibula autograft had similar functional outcome.     

Reconstruction of Acetabular Defects With Impaction Grafting in Primary Cemented Total Hip Arthroplasty Produces Favorable Results: Clinical and Radiographic Outcomes Over 6.4 Years on Average

BackgroundCemented primary total hip arthroplasty (THA) associated with acetabular reconstruction (AR) involving impacted bone grafting has been employed successfully in the revision of cavitary defects but the results are reportedly less predictable for segmental defects. The objective of the study is to evaluate the clinical and radiographic results of patients who had presented cavitary, segmental, and combined acetabular defects and received THA/AR involving impacted morselized cancellous bone autografts followed by rigorous postoperative management.MethodsClinical outcomes were assessed retrospectively of 154 patients who had been submitted to 169 THA/AR procedures performed by a single surgeon over a 15-year period. The Harris Hip Score system was applied to 103 patients, and the degree of acetabular migration was determined from radiograph images of 91 AR procedures, of which 40 were segmental/combined and 51 were cavitary reconstructions.ResultsThe frequency of aseptic acetabular loosening was 3% while that of thromboembolic events was 4.1%. According to Harris Hip Score, 87.4% of outcomes were classified as excellent/good after an average follow-up period of 6.4 years irrespective of the type of acetabular defect. Horizontal and/or vertical acetabular migrations and changes in angular tilt were observed in all ARs, although the minor movements detected did not undermine the structural stability of the reconstructions or the functional capacity of patients.ConclusionThe described THA/AR technique can be used effectively in the reconstruction of segmental/combined and cavitary acetabular defects. The success of the technique as applied to segmental/combined defects was attributed in part to the rigorous rehabilitation protocol with temporary postoperative weight-bearing restriction.     

Functional recovery after peripheral nerve injury via sustained growth factor delivery from mineral-coated microparticles

The gold standard for treating peripheral nerve injuries that have large nerve gaps where the nerves cannot be directly sutured back together because it creates tension on the nerve, is to incorporate an autologous nerve graft. However, even with the incorporation of a nerve graft, generally patients only regain a small portion of function in limbs affected by the injury. Although, there has been some promising results using growth factors to induce more axon growth through the nerve graft, many of these previous therapies are limited in their ability to release growth factors in a sustained manner and tailor them to a desired time frame. The ideal drug delivery platform would deliver growth factors at therapeutic levels for enough time to grow axons the entire length of the nerve graft. We hypothesized that mineral coated microparticles(MCMs) would bind, stabilize and release biologically active glial cell-derived neurotrophic factor(GDNF) and nerve growth factor(NGF) in a sustained manner. Therefore, the objective of this study was to test the ability of MCMs releasing growth factors at the distal end of a 10 mm sciatic nerve graft, to induce axon growth through the nerve graft and restore hind limb function. After sciatic nerve grafting in Lewis rats, the hind limb function was tested weekly by measuring the angle of the ankle at toe lift-off while walking down a track. Twelve weeks after grafting, the grafts were harvested and myelinated axons were analyzed proximal to the graft, in the center of the graft, and distal to the graft. Under physiological conditions in vitro, the MCMs delivered a burst release of NGF and GDNF for 3 days followed by a sustained release for at least 22 days. In vivo, MCMs releasing NGF and GDNF at the distal end of sciatic nerve grafts resulted in significantly more myelinated axons extending distal to the graft when compared to rats that received nerve grafts without growth factor treatment. The rats with nerve grafts incorporated with MCMs releasing NGF and GDNF also showed significant improvement in hind limb function starting at 7 weeks postoperatively and continuing through 12 weeks postoperatively when compared to rats that received nerve grafts without growth factor treatment. In conclusion, MCMs released biologically active NGF and GDNF in a sustained manner, which significantly enhanced axon growth resulting in a significant improvement of hind limb function in rats. The animal experiments were approved by University of Wisconsin-Madison Animal Care and Use Committee(ACUC, protocol# M5958) on January 3, 2018.