In vitro characterization of self-assembled anterior cruciate ligament cell spheroids for ligament tissue engineering.

Tissue engineering of an anterior cruciate ligament (ACL) implant with functional enthesis requires site-directed seeding of different cell types on the same scaffold. Therefore, we studied the suitability of self-assembled three-dimensional spheroids generated by lapine ACL ligament fibroblasts for directed scaffold colonization. The spheroids were characterized in vitro during 14 days in static and 7 days in dynamic culture. Size maintenance of self-assembled spheroids, the vitality, the morphology and the expression pattern of the cells were monitored. Additionally, we analyzed the total sulfated glycosaminoglycan, collagen contents and the expression of the ligament components type I collagen, decorin and tenascin C on protein and for COL1A1, DCN and TNMD on gene level in the spheroids. Subsequently, the cell colonization of polylactide-co-caprolactone [P(LA-CL)] and polydioxanone (PDS) polymer scaffolds was assessed in response to a directed, spheroid-based seeding technique. ACL cells were able to self-assemble spheroids and survive over 14 days. The spheroids decreased in size but not in cellularity depending on the culture time and maintained or even increased their differentiation state. The area of P[LA-CL] scaffolds, colonized after 14 days by the cells of one spheroid, was in average 4.57 ± 2.3 mm(2). Scaffolds consisting of the polymer P[LA-CL] were more suitable for colonization by spheroids than PDS embroideries. We conclude that ACL cell spheroids are suitable as site-directed seeding strategy for scaffolds in ACL tissue engineering approaches and recommend the use of freshly assembled spheroids for scaffold colonization, due to their balanced proliferation and differentiation.

[The French mesothelioma network from 1998 to 2013]. / Mésothéliome: les dispositifs en place en France « le réseau mésothéliome ¼ 1998-2013.

Mesothelioma is a rare disease less than 0.3% of cancers in France, very aggressive and resistant to the majority of conventional therapies. Asbestos exposure is nearly the only recognized cause of mesothelioma in men observed in 80% of case. In 1990, the projections based on mortality predicted a raise of incidence in mesothelioma for the next three decades. Nowadays, the diagnosis of this cancer is based on pathology, but the histological presentation frequently heterogeneous, is responsible for numerous pitfalls and major problems of early detection toward effective therapy. Facing such a diagnostic, epidemiological and medico-legal context, a national and international multidisciplinary network has been progressively set up in order to answer to epidemiological survey, translational or academic research questions. Moreover, in response to the action of the French Cancer Program (action 23.1) a network of pathologists was organized for expert pathological second opinion using a standardized procedure of certification for mesothelioma diagnosis. We describe the network organization and show the results during this last 15years period of time from 1998-2013. These results show the major impact on patient's management, and confirm the interest of this second opinion to provide accuracy of epidemiological data, quality of medico-legal acknowledgement and accuracy of clinical diagnostic for the benefit of patients. We also show the impact of these collaborative efforts for creating a high quality clinicobiological, epidemiological and therapeutic data collection for improvement of the knowledge of this dramatic disease.

Diffusion chamber system for testing of collagen-based cell migration barriers for separation of ligament enthesis zones in tissue-engineered ACL constructs.

A temporary barrier separating scaffold zones seeded with different cell types prevents faster growing cells from overgrowing co-cultured cells within the same construct. This barrier should allow sufficient nutrient diffusion through the scaffold. The aim of this study was to test the effect of two variants of collagen-based barriers on macromolecule diffusion, viability, and the spreading efficiency of primary ligament cells on embroidered scaffolds. Two collagen barriers, a thread consisting of a twisted film tape and a sponge, were integrated into embroidered poly(lactic-co-caprolactone) and polypropylene scaffolds, which had the dimension of lapine anterior cruciate ligaments (ACL). A diffusion chamber system was designed and established to monitor nutrient diffusion using fluorescein isothiocyanate-labeled dextran of different molecular weights (20, 40, 150, 500 kDa). Vitality of primary lapine ACL cells was tested at days 7 and 14 after seeding using fluorescein diacetate and ethidium bromide staining. Cell spreading on the scaffold surface was measured using histomorphometry. Nuclei staining of the cross-sectioned scaffolds revealed the penetration of ligament cells through both barrier types. The diffusion chamber was suitable to characterize the diffusivity of dextran molecules through embroidered scaffolds with or without integrated collagen barriers. The diffusion coefficients were generally significantly lower in scaffolds with barriers compared to those without barriers. No significant differences between diffusion coefficients of both barrier types were detected. Both barriers were cyto-compatible and prevented most of the ACL cells from crossing the barrier, whereby the collagen thread was easier to handle and allowed a higher rate of cell spreading.

Ultrastructural study of osteosarcomas.

The ultrastructural examination of four osteosarcomas (osteogenic, undifferentiated, and pleomorphic) is described. There are three types of tumor cells. Most of the cells are held in contact by desmosome-like tight junctions; they are atypical osteoblasts with cytoplasmic processes, dilated rough endoplasmic reticulum, mitochondria carrying calcific inclusions, lipid droplets surrounded by glycogen, and intracellular fine filamentous fibers. Other cells exhibiting varying degrees of osteoblastic maturity are also seen with multilobed nuclei, a clear cytoplasm, and straight bordered membranes. The last type is chondroid with abundant deposits of glycogen, lipid droplets, and undilated rough endoplasmic reticulum. The matrix is composed of fibrils without periodicity, scattered and deteriorated collagen fibers, and focal calcium deposits of hydroxyapatite crystals as in embryonal bone, dentine, or callus bone.

Treatment of tracheobronchial lesions by laser photoresection.

One hundred eleven patients underwent 205 laser photo-irradiation treatments using a Neodynium YAG laser with a flexible bronchoscope or an open tube for various tracheobronchial conditions, such as obstructing bronchogenic carcinomas, bronchial adenomas, and postintubation tracheal stenosis. The procedure was performed with either local or general anesthesia. Results were especially rewarding with endobronchial tumors. Tracheal stenoses were best treated by a combination of laser surgery and tracheal dilatation. No complication was encountered. Specific indications suggested for laser surgery are resection of inoperable tracheobronchial tumors, correction of tracheal stenosis, removal of surgical sutures, retrieval of tissue-embedded foreign bodies, and cauterization of hemorrhaging endobronchial tissues.

Benign tumors of the tracheobronchial tree. Endoscopic characteristics and role of laser resection.

We conducted a review of all the bronchoscopies performed at our institutions for benign tumors from 1980 to 1991 to determine the endoscopic characteristics of these lesions. We reviewed the charts, the endoscopic characteristics from our video records, and finally the pathologic findings of these cases. We tried to identify the effectiveness of laser resections in each group. Of a total of 3,937 patients, 185 (4.7%) were benign tumors. On these patients, 317 procedures were carried out. There were 3 myoblastomas, 53 papillomas, 1 adenoma, 8 chondromas, 4 fibromas, 45 hamartomas, 15 hamartochondromas, 6 lipomas, 19 angiomas, 5 leiomyomas, 4 schwannomas, 1 neurofibroma, and 21 amyloidomas. Results of laser resection were "very good" in 115 (62%) and "good" in 70 (38%). Complications were minimal: two mediastinal emphysemas, one pneumothorax, and one anesthesia-related cardiac arrest leading to the single death in this series. In general, benign tumors of the proximal endobronchial tree responded well to laser resection when their endoscopic appearance is recognized and prognosis known. Even when recurrent, repeated procedures can be performed easily with good results. This series is probably the largest in the world's literature about endoscopic recognition and the role of laser resection in patients presenting with benign endobronchial tumors.

Exsanguinating hemoptysis revealing the absence of left pulmonary artery in an adult.

Isolated absence of a pulmonary artery is an exceptional cause of massive hemoptysis. We report a 35-year-old woman with agenesis of the left pulmonary artery who presented with exsanguinating hemoptysis that prompted angiography with the aim to embolize the bleeding vessels selectively. The procedure could not be completed because of the presence of an anterior spinal artery branching from the aberrant systemic-to-pulmonary circulation. The patient successfully underwent an emergent pneumonectomy.

Primary intracerebral plasma cell granuloma: a light, immunocytochemical, and ultrastructural study of one case.

A case of primary intracerebral plasma cell granuloma without meningeal attachment is presented. Histologically, the lesion showed two different patterns. At the center, it displayed the features of a benign fibrous histiocytoma. At the periphery, a mixture of polyclonal plasma cells, lymphocytes, and "epithelial-like" cells was observed. Electron microscopy and immunocytochemistry confirmed the histiocytic nature of the "epithelial-like cells." The differential diagnosis and histogenesis of such a lesion are discussed.

Malignant transformation of an osteoblastoma of the skull: an exceptional occurrence. Case report.

What is apparently the first reported case of spontaneous malignant transformation of a benign osteoblastoma of the skull is described. The initial lesion was completely removed surgically and showed the histological features typical of a benign osteoblastoma. No radiotherapy was performed. Eleven years later the patient developed an osteosarcoma of the skull. Review of the literature showed that malignant transformation of benign osteoblastomas is extremely rare and could take place spontaneously. However, the risk of this occurring seems higher after inadequate initial treatment (curettage or partial excision). Follow-up monitoring of patients with osteoblastoma of the cranial vault is suggested.

[Detection of human papillomavirus by polymerase chain reaction in primary lung carcinoma]. / Détection de papillomavirus humain par amplification génique dans les cancers bronchiques primitifs.

Human papillomaviruses (HPV) have been implicated in the pathogenesis of human squamous cell carcinoma, specially of cervical carcinomas. In previous studies concerning primary lung cancer, DNA of HPV subtypes was detected by in situ hybridization or polymerase chain reaction (PCR), up to 30% of the cases, namely in squamous cell carcinomas. A series of 31 frozen biopsies of lung carcinomas (surgical biopsies or through fiber optic bronchoscopy) were examined for the presence of HPV DNA by nested PCR. Primers for the two steps were type-specific primers (6/11-16 and 18; kit Amplicis-HPV) for the transforming region of HPV. HPV-DNA was found in five tumors: in two of 18 cases of squamous cell carcinoma (11%), in one of four cases of adenocarcinoma, in one of six cases of small cell carcinomas and in the unic case of neuro-endocrin carcinoma. No case of the two large cell undifferentiated carcinomas was positive. There were three cases of HPV 6/11, one case of HPV 16, and one sample positive for HPV 6/11 and HPV 18. No morphologic changes consistent with HPV lesions were observed. The frequency of 11% among the squamous cell carcinomas is near those found by previous studies (9 to 20% for HPV 6-11-16-18). For the first time, HPVs have been detected in neuro-endocrin tumors, and this have to be confirmed by studies of many more cases. So HPV might play a role as promoter in carcinogenesis of any types of lung carcinoma, although at a low frequency.

[Pulmonary hypertension secondary to talc microemboli in a HIV seropositive heroin-addict woman]. / Hypertension artérielle pulmonaire secondaire à des micro-emboles de talc chez une héroïnomane séropositive au virus immunodépresseur humain.

Severe pulmonary hypertension presenting as acute cor pulmonare was observed in a HIV positive heroin addict. The usual aetiological investigations were negative. The apparently primary pulmonary hypertension was resistant oxygen and vasodilator therapy and was fatal in 6 months. Anatomopathological examination revealed the presence of talc microemboli in the pulmonary arterioles, severe medial hypertrophy and fibrous subendothelial thickening. The presence of the talc suggests that the pulmonary hypertension in this case was due to obstruction of the pulmonary vascular bed by the obstructive arterial lesions, despite the context of HIV infection.

[Pulmonary Wegener's disease mimicking tuberculosis]. / Maladie de Wegener pulmonaire simulant une tuberculose.

We report a case of early Wegener's disease mimicking tuberculosis in a 36 year old man. Lungs are solely affected for 6 months. Unusual appearance of radiographic features with large apical excavations explain the delai for the diagnosis. We discuss morphological features in the lungs and kidney.

[Bronchial mucoepidermoid carcinoma: apropos of 3 cases]. / Carcinome muco-épidermoïde bronchique: à propos de 3 observations.

We report three cases of bronchial mucoepidermoid carcinoma (BMEC) of low-grade malignancy with a relafase-free follow up. BMEC are rare tumors. The microscopic findings distinguish low-grade tumors which occur in children and young adults and high-grade tumors concerning older patients; this grading is based on the study of the epidermoid component. If possible, conservative therapy is appropriate in low-grade tumors. The prognosis of high-grade tumors is poor.

[Characteristic ultrastructural features of parathyroid gland cancer. A report on 9 cases (author's transl)]. / Particularités ultrastructurales du cancer des glandes parathyroïdes. A propos de 9 observations.

Authors have studied 9 parathyroid carcinomas under electron microscope (3% of their parathyroid lesions). Two types of the ultrastructural features were significant: 1) architectural: basal membrane abnormalities, sparse desmosomes, absence of nerve fiber; 2) cytological: nuclei atypism and abnormal mitochondria in tumors cells resembling dark chief cells. Also noticed were signs of cell hyperactivity and/or trouble in parathormon production or regulation.

[Prognostic value of UICC-1987 classification in primary lung cancers]. / Intérêt pronostique de la classification UICC-1987 des cancers bronchiques primitifs.

In order to test the validity of the new 1987-UICC classification (4th ed.) for lung cancer, data from 456 patients who underwent pulmonary resection for bronchogenic carcinoma from 1980 to 1985 were analysed retrospectively. Overall survival rate was 32.7% at 5 years. Prognostic significance of the tumour formula (pTNM) was assessed by multivariate analysis: N lymph node involvement appeared to be the most pejorative factor (p < or = 10(-5)), followed by the T factor (p < or = 0.002) and the M factor (p < or = 0.01). Respective survival curves were compared. Excluding the correlation between T1 and T2 tumours, the prognosis became significantly more unfavourable as the values for the pTNM classification criterion rose. Differences in prognosis between stages I and II, II and III A, III B and IV were not statistically significant for methodological reasons and inappropriate regroupings. Thus, our results partially confirm the prognostic value of the new international staging system. Meanwhile, this classification constitutes a prerequisite for the evaluation of end-results of therapy and the estimation of prognosis.

[Extrapancreatic hypoglycemic tumors. Apropos of a case of pleural fibrosarcoma]. / Tumeurs hypoglycémiantes extra-pancréatiques. Etude à propos d'un cas de fibrosarcome pleural.

The authors report a case of pleural fibrosarcoma associated with severe hypoglycaemia. Chromatography of the patient's serum and tumour fragments revealed a substance with a high molecular weight and an insulin-like activity. Professor Poffengarger's laboratory identified this substance as Non Suppressible Insulin-Like Protein or NSILAp which induces insulin-like effects on various tissues. A review of the literature is presented in the light of this case.

[Study of tracheobronchial healing after lung and heart-lung transplantation. Apropos of 64 anastomoses]. / Etude de la cicatrisation trachéo-bronchique après transplantations pulmonaires et cardio-pulmonaires. A propos de soixante-quatre anastomoses.

The authors report an analysis concerning the healing of tracheo-bronchial anastomoses after lung- and heart-lung transplantation. The present study includes 64 anastomoses selected from a total of 80 cases. Sixteen had to be excluded because of early post-operative death; none of these deaths was related to an air-way complication. Bronchial healing was assessed by bronchoscopic follow-up; the appearance of the suture-line was classified according to Couraud's grades. The initial reference was the examination at 2 weeks, which was compared to subsequent follow-up. At the initial assessment, 42 anastomoses were grade I, 4 were grade II, and 18 were grade III. The subsequent anatomic result was satisfactory for 52 sutures (81%). The complications were malacia in 2 cases, stenoses treated with a stenting device in 4 cases, dehiscence in 6 cases. The duration of ischemia and post-operative mechanical respiratory support, as well as the proximal or distal site of the anastomosis appeared to be of significant prognostic value.

[Induction treatment of locally advanced operable cancers of the esophagus. Prognostic significance of the histologic response]. / Traitement d'induction des cancers de l'oesophage opérables localement avancés. Signification pronostique de la réponse histologique.

OBJECTIVE: The purpose of this study was to determine the prognostic significance of the histopathological response to preoperative radio-chemotherapy in patients with locally advanced oesophageal cancer. METHODS: Among the 57 patients included in this open prospective study, the disease-free survival of 48 patients (8 females 40 males; mean age: 56.6 years +/- 8.4) who underwent an oesophagectomy after induction therapy for oesophageal squamous cell (n = 38) or adenocarcinoma (n = 10) was correlated with the histopathological findings. Chemoradiation included 2 cycles associating continuous 5 FU from D1 to 5 and from D22 to 26, cisplatyl on D1 and D22, 15 Gy/5d from D1 to 5 and from D22 to 26. Histopathological response was assessed on the operative specimens by routine examination of serial thin sections each 5 mm along the full oesophageal length, the resection margins and the lymph node dissection. RESULTS: A wide interindividual variability was seen regarding tissue changes related to induction therapy, with a grading in tumor regression and the possibility of dissociated effects on the various treatment targets: tumor, adenopathy and vessel invasion. The 5-year probability of disease-free survival was 22% for the 48 resected patients. The presence of a complete histopathological response (n = 12) did not preclude metastatic spread in half the cases. Furthermore, it did not result in improved survival when compared to that of non-responder patients. Survival of patients who had a complete or major oesophageal response (n = 29, 35% at 5 years) was significantly lower than that of patients who were operated on during the same period for a superficial oesophageal cancer at presentation (n = 29, 57% at 5 years; P = 0.03). After multivariate analysis according to the Cox model, downstaging of the primary tumor was not identified as an independent predictor of disease-free survival. CONCLUSIONS: Pathologic assessment of tumor regression on the operative specimen provides little prognostic information.

[Endosonography in the preoperative evaluation of cancers of the esophagus]. / L'endosonographie dans l'évaluation pré-opératoire des cancers de l'oesophage.

OBJECTIVE: To assess the diagnostic accuracy of endoscopic ultrasonography (EUS) for the local and regional staging of esophageal cancer, and its possible alteration resulting from the performance of preoperative chemoradiation. METHODS: Prospective study of 85 consecutive patients with esophageal cancer evaluated by EUS and operated on between January 1992 and December 1995. 28 of these patients had received previous induction therapy. In all cases, EUS examination was performed by the same physician not informed about the results of previous morphological explorations. Histopathological analysis of all operative specimens was performed by the same pathologist, not informed about the results of EUS. Data were collected by another independent observer. RESULTS: EUS examination resulted in incomplete staging in 8 patients (9.5%) with severe stenosis precluding endoscope passage. The accuracy, specificity and sensitivity of EUS in detecting the depth of esophageal involvement (T0-2 vs. T3-4) were 82.3%, 78%, and 86% respectively, and 72%, 70%, and 73% respectively for lymph node metastasis. The overall accuracy of EUS in identifying the preoperative stage was 67%, with a clear-cut alteration when patients had received induction therapy (61% vs 72%). On the other hand, 7 (64%) of the 11 patients thought to have a complete response at endosonography had no residual tumor. CONCLUSION: EUS provides precise information for the preoperative identification of locally advanced esophageal tumor, even after induction therapy. The latter alters the diagnostic accuracy of EUS, although complete responders could be identified in two-thirds of cases.

[Tracheobronchial healing after lung and heart-lung transplantations. Apropos of 64 anastomoses]. / Etude de la cicatrisation trachéo-bronchique après transplantations pulmonaires et cardio-pulmonaires. A propos de 64 anastomoses.

The authors report an analysis concerning the healing of tracheo-bronchial anastomoses after lung- and heart-lung transplantation. The present study includes 64 anastomoses selected from a total of 80 cases. Sixteen had to be excluded because of early post-operative death; none of these deaths was related to an air-way complication. Bronchial healing was assessed by bronchoscopic follow-up; the appearance of the suture-line was classified according to Couraud's grades. The initial reference was the examination at 2 weeks, which was compared to subsequent follow-up. At the initial assessment, 42 anastomoses were grade 1, 4 were grade II, and 18 were grade III. The subsequent anatomic result was satisfactory for 52 sutures (81%). The complications were malacia in 2 cases, stenoses treated with a stenting device in 4 cases, dehiscence in 6 cases. The duration of ischemia and postoperative mechanical respiratory support, as well as the proximal or distal site of the anastomosis appeared to be of significant prognostic value.