Regeneration pattern of cardiac and skeletal muscle after transplantation into a skeletal muscle bed in rats

Background: The ability of skeletal muscle to regenerate after injury is well established. In contrast, cardiac muscle is incapable of regeneration and recovery after injury. The aim of the present study was to evaluate and compare the regeneration pattern of cardiac and skeletal muscle after transplantation into a skeletal muscle bed in rats. Methods: The following group of transplants were performed at the site prepared by removing the host extensor digitorum longus (EDL) muscle. The first group consisted of cardiac muscle transplanted as one piece or after mincing into 1-mm pieces. The second group included cotransplants of cardiac and skeletal muscle minces that were intermixed. Entire EDL muscle or minced EDL muscle were also transplanted for comparison. Rats were sacrificed 3–30 days after transplantation for morphological analysis. Results: The results demonstrated that skeletal muscle transplants underwent rapid regeneration, and by 30 days the entire muscle was filled with regenerated myofibers. In transplants of cardiac muscle significant inflammation, myocardial degeneration and necrosis were observed. In spite of the necrosis and fibrosis, the presence of a few regenerated myotubes in the outer region was observed. In cardiac and skeletal muscle cotransplants, the inflammation was restricted to cardiac tissue; however, by 30 days the entire contransplant was filled with regenerated myotubes and myofibers. Conclusions: These results show that skeletal muscle is capable of growth, regeneration, and integration with the cardiac muscle after cotransplantation. Combination of skeletal and cardiac muscle may prove useful in defining the cellular processes necessary for enhancing cardiac repair after injury. © 1995 Wiley-Liss, Inc.     

Mineralization and calcium fixation within a porous apatitic ceramic material after implantation in the femur of rabbits

Cylinders (0.8 cm long, 1.0 cm of diameter and with an axial hole), constituted, after firing, of a ceramic mixture of hydroxylapatite (HA) and beta-tricalciumphosphate (beta-TCP) in a 10:1 ratio, were implanted into mid-diaphyseal defects of one femur of 20 rabbits and stabilized with intramedullary rods. The implantation sites were checked radiographically every month, and after 3 months (3 animals) and 6 months (17 animals) the rabbits were sacrificed and the implants with the surrounding tissue were embedded in methylmethacrylate, cut to thick sections and analyzed by scanning electron microscopy (SEM). Porosimetric and x-rays diffraction analyses were carried out before and after implantation of the cylinders, and the state of mineralization at the bone-implant interface was determined by EDAX microprobe analysis. Bony callus formation started at 1 month at the osteotomy sites, as judged by radiography, but after 3 months a not-mineralized zone had still been demonstrated between bone and the implants. At 6 months, 13 implants showed themselves firmly fixed in their implantations beds, while 4 implants were only incorporated at their proximal ends. In bone contact zones, an enrichment of Ca2+ was displayed by microanalytical techniques in the outer zone of the implanted samples which may be explained by an apparent additional phase transformation of HA into TCP, thanks to the change of the Ca/P ratio, that takes place in vivo.     

Regeneration of the junctional epithelium and connective tissue after transplantation of detergent-processed allo-teeth

The authors have developed a new artificial dental implant and evaluated it in a dog model in terms of its potential to produce: I) regeneration of junctional epithelium; II) regeneration and attachment of connective tissue. The implants were constructed from allo-teeth. We removed the cell components from the periodontal ligaments of these teeth with a detergent (1% TritonX-100); the remaining acellular periodontal ligament acted as an extracellular matrix upon which regeneration and attachment could proceed. We placed 10 of these implants in the just-extracted sites of three beagle dogs. We observed regeneration of both junctional epithelium and connective tissue at all implant sites after 3 months. The connective tissue was attached in all cases. Use of the acellular periodontal ligament as an extracellular matrix may facilitate regeneration of host periodontal ligament tissue, thus contributing to recovery of host immunological defense and long-term oral function.     

Identification of wear particles in tissue after implantation of different plastic materials.

A method is shown by which to identify wear particles of implanted plastic materials in tissue. A Mettler FP2 hot stage was used to determine the melting point of different elastomer particles in normal histological sections between the crossed nicols. By using a microcolorimeter Type MPE (Leitz, Wetzlar, Germany), additional information about the molecular degradation of wear particles was obtained.     

Study of the healing process after transplantation of pasteurized bone grafts in rabbits

Different bone allografts (pasteurized, autoclaved, and frozen) were compared based on their osteoinductive properties. Our primary purpose was to examine the biologic qualities of pasteurized allografts, as pasteurization inactivates most viruses transmitted by transplantation. Frozen, pasteurized, and autoclaved allografts were packed into a standard defect of rabbit ulna. The animals were sacrificed at 2 and 4 weeks after surgery. The parts of bones with experimental defects were explored en bloc, and a roentgenogram was carried out. Ulna bone samples were then embedded in methyl-methacrylate. Roentgenograms showed that after 2 weeks, calluses were well-formed, but irregular in shape in all 3 types of allografts. After 4 weeks, the calluses were regular in shape in all but the autoclaved grafts. After 2 weeks, the healing processes had begun in the frozen and pasteurized grafts, with the reaching approximately the same stage, while in the autoclaved grafts these processes were not seen and the bone particles were surrounded by connective tissue without any changes. After 4 weeks, osteoinductive processes were very strong, with the first signs of complete bone remodeling at the bone edges of the defect in pasteurized and frozen allografts. The osteoinductive values of these 2 types were very high and similar. Autoclaved allografts, on the other hand, had very low osteoinductive values, as they were still at the very beginning of the healing process. Histomorphometric analysis revealed a significant difference in both newly formed osteoid thickness and osteoblast number per microm of bone surface in all experimental groups (P < 0.005). Values of osteoid thickness and osteoblast number were significantly higher in both frozen and pasteurized grafts when compared with the autoclaved ones (P < 0.005). Osteogenic properties of pasteurized bone allografts were preserved, and the allografts have been gradually replaced with newly formed bone. As such, pasteurized bone grafts from a bone bank have approximately the same biologic validity as frozen grafts, while autoclaved grafts impair bone healing.     

Regeneration of the liver in mice treated with a mixture of hepatotoxins in delayed periods after bone marrow transplantation

Possible effect of bone marrow cells on liver regeneration was studied in mice injected with a mixture of hepatotoxins (allyl alcohol and CCl₄) in a dose equal to LD₅₀. The mixture of hepatotoxins was used to minimize the restitution regeneration of the liver. The dose of allyl alcohol causing (in combination with CCl₄) maximum liver damage was selected beforehand. Increasing the dose of allyl alcohol in the two-component mixture resulted in more severe necrosis of the liver. The maximum dose of alcohol (50 mg/kg) in combination with CCl₄ caused irreversible injury to the liver leading to 100% mortality after 2–4 days. In radiation chimeras reconstituted by bone marrow cell transplantation, in which liver damage was induced by a mixture of hepatotoxins containing the maximum dose of allyl alcohol, we observed normalization of liver tissue structure and function. The mechanism of this effect is not clear. __________ Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 141, No. 4, pp. 462–465, April, 2006     

新西兰兔下颌骨牵引成骨早期拆除牵引装置植入钛钉对新骨形成的影响

目的：利用锥形束CT（ CBCT）研究新西兰兔下颌骨矩形截骨后,早期拆除牵引装置植入钛钉对新骨形成 的影响,探讨牵引成骨早期拆除牵引装置的可能性。方法：新西兰兔下颌骨矩形截骨12 mm×4 mm,植入自制牵 引装置。牵引5 d 后,随机分为稳定4 周、稳定8 周、钛钉植入3 组。前2 组分别于稳定期4 周、8 周时处死动物;钛 钉植入组,稳定期4 周时,取出牵引器植入钛钉,4 周后处死动物;拍摄CBCT和X线片。结果：兔下颌骨垂直高 度平均增加（2.25±0.41）mm。稳定4 周、8 周和钛钉植入组的术区CT值分别为（80.00±12.36）Hu、（293.00±37.66） Hu和（289.25±38.87）Hu。稳定4 周术区CT值低于稳定8 周和钛钉植入组,后2 组CT值相近。结论：早期拆除 牵引装置植入钛钉与常规固定相比,骨形成没有明显影响,临床应用可有效地缩短牵引成骨的时间。     

Evaluation of a hybrid scaffold/cell construct in repair of high-load-bearing osteochondral defects in rabbits

The objective of this study was to evaluate the feasibility and potential of a hybrid scaffold system in large- and high-load-bearing osteochondral defects repair. The implants were made of medical-grade PCL (mPCL) for the bone compartment whereas fibrin glue was used for the cartilage part. Both matrices were seeded with allogenic bone marrow-derived mesenchymal cells (BMSC) and implanted in the defect (4 mm diameter x 5 mm depth) on medial femoral condyle of adult New Zealand White rabbits. Empty scaffolds were used at the control side. Cell survival was tracked via fluorescent labeling. The regeneration process was evaluated by several techniques at 3 and 6 months post-implantation. Mature trabecular bone regularly formed in the mPCL scaffold at both 3 and 6 months post-operation. Micro-Computed Tomography showed progression of mineralization from the host-tissue interface towards the inner region of the grafts. At 3 months time point, the specimens showed good cartilage repair. In contrast, the majority of 6 months specimens revealed poor remodeling and fissured integration with host cartilage while other samples could maintain good cartilage appearance. In vivo viability of the transplanted cells was demonstrated for the duration of 5 weeks. The results demonstrated that mPCL scaffold is a potential matrix for osteochondral bone regeneration and that fibrin glue does not inherit the physical properties to allow for cartilage regeneration in a large and high-load-bearing defect site.     

Different inflammatory response and oxidative stress in neointimal hyperplasia after balloon angioplasty and stent implantation in cholesterol-fed rabbits

Inflammatory responses appear to play an important role in the occurrence of restenosis following coronary intervention. However, the contribution of C-reactive protein (CRP) and oxidative stress to restenosis after balloon angioplasty and stent implantation remains unclear. The aim of this study was to examine this issue using hyperlipidemic rabbits. Rabbits were divided into two groups; they were fed with a 0.5% cholesterol diet and with a mixed 0.5% cholesterol and 0.5% probucol diet. Each group of rabbits underwent balloon injury and stent implantation in right and left iliac arteries, respectively. Eight weeks after the intervention, we examined luminal stenosis, neointimal hyperplasia, immunoreactivity for macrophage, CRP and oxidized phosphatidylcholine (oxPC), and also the expression of CRP mRNA. The degrees of neointimal hyperplasia and immunopositive areas (%) for macrophage, CRP, and oxPC in the neointima were significantly higher after stent implantation than after balloon injury, but CRP mRNA was undetectable in either artery. Anti-oxidant probucol reduced angiographic stenosis, neointimal hyperplasia, and macrophage- and oxPC-positive areas much more significantly after stenting. The results demonstrate that the inflammatory response to the development of neointimal hyperplasia differs after balloon injury and stent implantation and that CRP deposition and oxidative stress might be involved more significantly in neointimal development after stent implantation.     

Imaging and morphology of the paracondylar process in a dry adult human skull: a case report

An unusually long paracondylar process (PCP) was found articulating with the transverse atlantal process in a dry adult male skull. It was 13 mm long and 19 mm in circumference at its middle and projected to a lower plane than the styloid process of the same side. A radiograph revealed the structure of spongiosa of the PCP to be like mastoid cells. PCP can lead to clinical symptoms due to alterations in posture and functional limitations in neck movements. Thus awareness of this process, its topographical relations and its attendant problems is of importance to clinicians, radiologists, surgeons and chiropractors. In symptomatic cases surgical resection of this process can be attempted to relieve the symptoms. The craniovertebral junction constitutes a unique part of the somite-derived axial skeleton; the lowest part of the basioccipital component including this process develops from occipital sclerotomes. Developmental variations in this region can resemble deformities.     

Restricted chimerism of peripheral blood cells after transplantation of allogeneic hemopoietic tissue in rabbits.

Allogeneic bone marrow or spleen cells were transplanted in rabbits treated with a single dose of 110 kmg/g of cyclophosphamide or four doses of 50 mg/kg. Chimerism of peripheral blood cells occurred in about 50% of the recipients, lasting from about the 14th day until 180 days after transplantation. The chimerism was limited to some kinds of cells since dissociation of proliferation of hemopoietic cells and split chimerism of lymphocytes were observed. Dissociation of proliferation of hemopoietic cells consisted in the appearance of only one cell line, namely lymphocytes, whereas the erythrocyte cells did not proliferate. Split chimerism occurred within lymphocytes, manifested by the presence of the donor's T lymphocytes but absence of proliferation of the donor's B lymphocytes.     

Enlarged parietal foramina: a rare finding in a female Greek skull with unusual multiple Wormian bones and a rich parietal vascular network

Enlarged parietal foramina (>5 mm) is an extremely rare developmental defect of the parietal bone, which is distinguished from the normal small parietal foramina, as genes associated with this entity have been identified, suggesting that it is hereditary in nature. We describe a dry skull of a 35-year-old female, with enlarged parietal foramina symmetrically situated bilaterally, oval in shape, measuring 4.5 × 9.3 mm (right) and 4.9 × 9.2 mm (left) in size. The foramina coexisted with multiple Wormian bones in several sites of the skull. On the inner parietal bone surface, the anterior, posterior and lateral foramina’s rims carried grooves, which were continuous with the middle meningeal vessels’ branches, indicating that a rich vascular network existed around the foramina. These vascular grooves also notched the external table at the margin of the foramina, which suggests a potential communication between the meningeal and the scalp vessels. In addition, this vascular variation should be taken into consideration when performing surgical interventions in the area, because the large vascular supply to the foramina is a possible source of extensive bleeding. Moreover, the interaction of intracranial and extracranial veins and the fact that the blood flows in them in both directions, as they are valveless, could represent a possible pathway for infections to spread in the cranial cavity.     

Comparison of two commercial methods for measurement of cytomegalovirus load in blood samples after renal transplantation

A cohort of 77 renal transplant recipients was prospectively studied for comparison of two commercially available cytomegalovirus (CMV) load assays, i.e., the COBAS AMPLICOR CMV Monitor test (Amplicor), using plasma samples, and the Murex Hybrid Capture System (HCS), using whole blood. The manufacturer of the HCS assay changed the version of the test from 1.0 (HCS-1) to 2.0 (HCS-2) after the first 37 patients had been tested. Despite the differences in principle and type of specimen used, the Amplicor and HCS assays gave comparable results. The regression line correlating the HCS-1 assay to the Amplicor assay was similar to that correlating the HCS-2 assay to the Amplicor assay. The HCS results could be converted to Amplicor-equivalent units by using linear-regression equations [log(10) HCS-1 result = 0.49 (log(10) Amplicor result) + 2.58, and log(10) HCS-2 result = 0.61 (log(10) Amplicor result) + 2.18]. The HCS-2 assay appeared to have the lowest detection limit, followed by the Amplicor assay and then the HCS-1 assay. When a sliding scale of cutoff values in Amplicor-equivalent units (>1,000, >2,500, >6,000, >16,000, >40,000, and >100,000 copies/ml) was applied to diagnose CMV disease, similar patterns of sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were observed with the Amplicor and HCS assays. A cutoff value of >40,000 copies/ml has a low sensitivity (Amplicor, 29.4%; HCS, 41.2%) but is specific (Amplicor, 96.7%; HCS, 93.3%) and can be used for the differential diagnosis of CMV disease (PPV, 71.4% [Amplicor] or 63. 6% [HCS]; NPV, 82.9% [Amplicor] or 84.8% [HCS]). A lower cutoff value of >1,000 copies/ml improves the sensitivity (Amplicor, 76.5%; HCS, 82.4%) and has a high NPV (Amplicor, 91.8%; HCS, 94.2%) but, due to the low PPV (Amplicor, 46.2%; HCS, 56%), is useful only for exclusion of CMV disease.