Influence of cyclic bending loading on in vivo skeletal tissue regeneration from periosteal origin.

INTRODUCTION: Periosteum osteogenic and chondrogenic properties stimulate the proliferation then differentiation of mesenchymal precursor cells originating from its deeper layers and from neighboring host tissues. The local mechanical environment plays a role in regulating this differentiation of cells into lineages involved in the skeletal regeneration process. HYPOTHESIS: The aim of this experimental animal study is to explore the influence of cyclic high amplitude bending-loading on skeletal tissue regeneration. The hypothesis is that this mechanical loading modality can orient the skeletogenesis process towards the development of anatomical and histological articular structures. MATERIAL AND METHODS: A vascularised periosteal flap was transferred in close proximity to each knee joint line in 17 rabbits. On one side, the tibiofemoral joint space was bridged and loading occurred when the animal bent its knee during spontaneous locomotion. On the other side, the flap was placed 12 mm distal to the joint line producing no loading during bending. Tissue regeneration was chronologically analyzed on histologic samples taken from the 4th day to the 6th month. RESULTS: The structure and mechanical behavior of regenerating tissue evolved over time. As a result of the cyclic bending-loading regimen, cartilage tissue was maintained in specific areas of the regenerating tissue. When loading was discontinued, final osteogenic and fibrogenic differentiation occurred in the neoformed cartilage. Fissures developed in the cartilage aggregates resulting in pseudo-gaps suggesting similar processes to embryonic articular development. Ongoing mesenchymal stem cells stimulation was identified in the host tissues contiguous to the periosteal transfer. DISCUSSION: These results suggest that the pseudarthrosis concept should be reconsidered within the context of motion induced articular histogenesis.

Intraoperative immunophotodetection for radical resection of cancers: evaluation in an experimental model.

The aim of our study was to assess the technique of immunophotodetection (IPD) in intraoperative situations in an experimental model and to determine its capacity to detect very small tumor masses. IPD is a recent technology involving fluorescent dye-labeled monoclonal antibodies (MAbs) directed against tumor-associated antigens. Up to now, no intraoperative device for IPD has been developed, and limits of detection of the technique are unknown. MAb-dye conjugates were prepared using the anti-carcinoembryonic antigen MAb 35A7 labeled with indocyanine and (125)I. Time-dependent (6, 12, 24, 48, and 96 h post i.v. injection) and dose-dependent (10, 40, and 100 microg of conjugate) biodistribution studies were performed in nude mice bearing an LS174T peritoneal carcinomatosis demonstrating high tumor uptake (up to 21% of the injected dose/g of tumor 48 h postinjection). Intraoperative IPD was studied, using a newly developed device, in 16 mice 48 h after i.v. injection of 40 microg of the (125)I-MAb 35A7-indocyanine conjugate. The fluorescent status of 333 biopsies was compared with their histological analysis. Sensitivity was 90.7%, specificity was 97.2%, the positive predictive value was 94.7%, and the negative predictive value was 94.9%. Detection of very small nodules (<1 mg in weight or <1 mm in diameter) was possible. However, we observed a decrease in sensitivity as a function of tumor mass: 100% for nodules >10 mg versus 78% for nodules < or =1 mg. These experiments demonstrate that intraoperative IPD is easy to use and associated with high sensitivity and specificity, even for low tumor masses. On the basis of these encouraging results, intraoperative IPD should be assessed in a clinical study.

Resorption kinetics of eggshell: an in vivo study.

Eggshell has been recently introduced as a bone substitute candidate in reconstructive surgery. The aim of this experimental study study was to determine its degradation rate in both a skeletal and extraskeletal site. In experiment 1, eggshell particles with four different sizes (50, 75, 150, and 300 microns in diameter) were implanted in subcutaneous pouches of 30 rats. In experiment 2, a fragment of ostrich eggshell was implanted on the nasal dorsum of 10 rats. Animals were sacrificed at 1 (N = 10), 2 (N = 10), and 4 months (N = 10) during the first stage of the study, and at 1 year during the second stage of the study. The results were assessed by X-ray examination and routine histological techniques. In experiment 1, all animals healed uneventfully. At 1 month, only 50-micron particles had undergone resorption. At 2 months, both 50- and 75-micron particles had undergone resorption. At 4 months, the 150- and 300-micron particles were resorbed incompletely. Histologically, the eggshell elicited a mild inflammatory reaction at 1 month that decreased progressively at further stages. In experiment 2, all animals except one healed uneventfully. Radiologically, the eggshell implant displayed a noticeable stability. Histologically, seven of nine implants were encapsulated, but two of them were surrounded by a bony rim. In conclusion, eggshell is a resorbable implant, but the degradation kinetic is size dependent. Large ostrich grafts are also suitable as onlay graft, but a complementary osteosynthesis is recommended to enhance osteointegration.

Comparative study of three different membranes for guided bone regeneration of rat cranial defects.

The purpose of the study was to compare the efficacy of a non-resorbable polytetrafluoroethylene (e-PTFE) membrane (Gore-Tex) versus a resorbable polyglactin membrane (Vicryl) and a newly designed collagenic membrane for enhancing bone regeneration on rat skull defects. The study was conducted on 30 adult Wistar rats. On each animal, two symmetrical, 6 mm wide, full-thickness, skull defects were created in the parietal regions. The right defect was chosen as the experimental site and the left one was left empty as a control. Each experimental site was covered by an inner and outer membrane. The 30 rats were divided into three groups: In group 1 (n=10), a non-resorbable polytetrafluoroethylene (e-PTFE) membrane was used. In group 2 (n= 10), a resorbable polyglactin 910 membrane was used. In group 3 (n= 10), a collagen membrane processed from avian eggshell was used. In each group, the animals were euthanized at 60 days. The harvested specimens were processed for contact radiography and standard histological examination. The results were assessed by a Fisher's exact test. In group 1, partial bone healing was observed in seven out of 10 animals and complete in three out of 10 animals (P<0.001). In group 2, no or minimal bone healing was observed in seven out of 10 animals and partial bone healing was observed in three out of 10 animals. In group 3, no or minimal bone healing was observed in nine out of 10 cases and partial bone healing in only one animal. In conclusion, only the non-resorbable e-PTFE membrane group exhibited a favourable result in this study. This study suggests that the structure of the membrane is at least as important as its composition.

Comparison of pericranium and eggshell as space fillers used in combination with guided bone regeneration: an experimental study.

PURPOSE: The purpose of the study was to determine the potential adjunctive role of 2 space fillers, used in combination with guided tissue regeneration, on bone regeneration in rat skull defects. MATERIALS AND METHODS: The study was conducted on 45 adult Wistar rats. A bilateral 6-mm-wide full-thickness skull defect was created in the parietal region of each animal. The right defect was chosen as the experiment site, and the left defect was left empty as a control. Each experiment site was covered by an inner and outer polytetrafluoroethylene membrane. The 45 rats were divided into 3 groups; no space filler between the 2 membranes was used in group I (n = 15), a free pericranial autograft was used as a space filler between the 2 membranes in group II (n = 15), and purified eggshell powder was used as space filler between the 2 membranes in group III (n = 15). Five animals in each group were killed at 15, 30, and 90 days. The harvested specimens were subjected to contact radiography and standard microscopic examination, and the rates of osteogenesis were assessed by a semiquantitative method. RESULTS: No evidence of bone regeneration was seen in any animals of the three groups at 15 days. At 30 days, bone regeneration only appeared in group I (P > .05). At 90 days, complete bone regeneration was observed in the group I in 3 of 5 animals (P < .05). In group II, the pericranial graft showed no osteogenic properties. In group III, the eggshell powder showed no resorption, but no osteoconduction was noticed. CONCLUSIONS: Although the osteogenic mechanism of guided tissue regeneration is not clear, this study suggests that the physical properties of the membrane are more important than the use of an adjunctive space filler.

Involvement of circulating CEA in liver metastases from colorectal cancers re-examined in a new experimental model.

Both experimental and clinical data show evidence of a correlation between elevated blood levels of carcinoembryonic antigen (CEA) and the development of liver metastases from colorectal carcinomas. However, a cause-effect relationship between these two observations has not been demonstrated. For this reason, we developed a new experimental model to evaluate the possible role of circulating CEA in the facilitation of liver metastases. A CEA-negative subclone from the human colon carcinoma cell line CO115 was transfected either with CEA-cDNA truncated at its 3' end by the deletion of 78 base pairs leading to the synthesis of a secreted form of CEA or with a full-length CEA-cDNA leading to the synthesis of the entire CEA molecule linked to the cell surface by a GPI anchor. Transfectants were selected either for their high CEA secretion (clone CO115-2C2 secreting up to 13 microg CEA per 10(6) cells within 72 h) or for their high CEA membrane expression (clone CO115-5F12 expressing up to 1 x 10(6) CEA molecules per cell). When grafted subcutaneously, CO115-2C2 cells gave rise to circulating CEA levels that were directly related to the tumour volume (from 100 to 1000 ng ml(-1) for tumours ranging from 100 to 1000 mm3), whereas no circulating CEA was detectable in CO115 and CO115-5F12 tumour-bearing mice. Three series of nude mice bearing a subcutaneous xenograft from either clone CO115-2C2 or the CO115-5F12 transfectant, or an untransfected CO115 xenograft, were further challenged for induction of experimental liver metastases by intrasplenic injection of three different CEA-expressing human colorectal carcinoma cell lines (LoVo, LS174T or CO112). The number and size of the liver metastases were shown to be independent of the circulating CEA levels induced by the subcutaneous CEA secreting clone (CO115-2C2), but they were directly related to the metastatic properties of the intrasplenically injected tumour cells.

The effect of pentosan polysulphate on bone healing of rat cranial defects.

The purpose of the study was to determine the efficacy of pentosan polysulphate, used in combination with guided bone regeneration on rat skull defects. The study was conducted on 45 adult Wistar rats. On each animal two symmetrical 6 mm wide, full-thickness, skull defects were created in the parietal regions. The right defect was chosen as the experimental site and the left one was left empty to provide a control. Each experimental site was covered by an inner and outer polytetrafluoroethylene membrane. The 45 rats were divided into 3 groups: in group I (n = 15), carboxymethyl cellulose, used as a delivery vehicle, was injected between the two membranes; in group II (n = 15), 1 mg of pentosan polysulfate was added to the carboxymethyl cellulose vehicle; in group III (n = 15), purified micronized eggshell powder was added to the mixture of pentosan polysulfate and carboxymethyl cellulose between the two membranes. In each group, the animals were sacrificed at 42 days. The harvested specimens were processed for contact radiography and standard histological examination. The results were assessed by a Fisher's exact test. All animals, except one, healed uneventfully. In group I, partial bone healing was observed in 14 out of 15 animals. In group II, partial bone healing was observed in 13 out of 15 animals, and complete bone healing in 1 out of 15 cases. In group III, partial resorption of the eggshell implant was observed with a partial bone healing in only 2 cases (P < 0.001). In conclusion, significant bone regeneration was observed with the membranes alone. The use of pentosane polysulphate did not result in additional bone gain. The use of particulate material as a space maintainer is also questionable.

[Cancer of the breast: the return of lymphoscintigraphy?]. / Cancer du sein: le retour de la lymphoscintigraphie?

Lymphoscintigraphy, after arousing great hope in the past in the field of breast cancer, has now been abandoned. The inability of this examination to predict the metastatic status of the nodes, and progress in therapeutic concepts have led to abandoning this technique. However, certain problems encountered by regional irradiation programmes and the work concerning sentinel node detection may bring this technique back into the spotlight. Lymphoscintigraphy may make it possible to adopt an individual approach, case by case, of the lymphatic drainage basins in breast tumors, thus enabling certain patients to benefit from regional irradiation when it would not have been traditionally recommended for this irradiation. Another aspect concerns the problem of the volumes irradiated. Work carried out with lymphoscintigraphy has enabled internal mammary chain nodes to be precisely located. Theses studies show the necessity of adapting the irradiation field to each individual case, but the clinical impact is limited, in the end, by the low recurrence rate in the internal mammary chain area. However, the new techniques of computer merging of scintigraphic and scanner images could enable the spatial position of the nodes in the upper axillary and supraclavicular regions to be determined. This would have, a priori, much wider clinical impact. Lymphoscintigraphic detection of the sentinel node is another field of major interest, but this technique is in competition with staining techniques. This procedure leads to a large reduction in morbidity of axillary surgery in 70% of patients. The use of techniques for detecting micrometastases in the sentinel node opens prospects in terms of prognosis. The qualities of differents radiotracers and different injection sites possible are also discussed.

Experimental assessment of the risk of tumor recurrence after laparoscopic surgery.

BACKGROUND: The aim of this experimental study was to evaluate the risk of tumor recurrence after laparoscopic cecal resection (LCR) of colonic carcinoma in the rat. METHODS: The experimental cancer consisted of one million cells (DHK/K12), incorporated in an extracellular matrix, placed and secured to the cecal serosa in 110 BD9 rats. Four weeks later, all animals were reoperated through a laparotomy to control tumor growth, and animals with diffuse carcinomatosis were excluded. Eligible animals were randomized either to laparoscopic cecal resection (group LCR, n = 10), to open resection (group OCR, n = 13), or to a control group without resection (group C, n = 13). Resection was always considered as macrocopically complete. All animals were killed 4 weeks after the resection to determine the tumor recurrence and quantify carcinomatosis. RESULTS: We noted diffuse carcinomatosis in 70% of rats in groups C and LCR versus 23% in group OCR (p = 0.038). For tumors noted as S- (not extending outside the serosa), diffuse carcinomatosis was observed in all animals of group C (3 of 3), in 6 of 8 in group LCR, and 0 of 6 in group OCR (p = 0.004). The rate of port site or incisional metastases was not significantly different between groups. CONCLUSIONS: These preliminary results demonstrated the deleterious impact of the laparoscopy for resection of large bowel malignancy. LCR increased significantly the incidence of a diffuse carcinomatosis even when performed for locally noninvasive tumors (S-).

Powdered eggshell: a pilot study on a new bone substitute for use in maxillofacial surgery.

The present study is a preliminary report on the use of hen's eggshell as a possible bone substitute. In the first part of the study, particles ranging from 400 microns to 600 microns in diameter were bioassayed in an intramuscular pouch in rodents. This material was found to be biocompatible, but appeared not to have osteoinductive capacities. In the second and third part of the study, this material was used as an interpositional graft material in critical-size defects of rat mandibles and rabbit skulls. At 2 months, a morphologic restoration was obtained using the graft, but the healing was only achieved by fibrous union. In the fourth part of the study, the material was experimented on as an onlay bone graft on rabbit mandibles. A 6-month follow-up of the implant confirmed its stability. In conclusion, the use of this safe and inexpensive material is suggested for filling limited bone defects in non-weight-bearing areas. The use of eggshell powder for bone augmentation may also be considered, after further studies, to assess its long-term stability.