Effect of nerve growth factor on cultured human chondrocytes.

Nerve growth factor (NGF) is involved in several joint diseases. It participates in nociception and neurogenic inflammation and its concentrations increase in synovial fluid and tissue from arthritis. However, data about its role in articular cartilage are scant and conflicting. This study analysed effects of different NGF concentrations on cultured human chondrocytes by evaluating cell proliferation, cell phenotype, and gene expression. The MTT test excluded an influence on cell viability. Alcian blue and S100 staining demonstrated that NGF may induce de-differentiation of the chondrocyte phenotype. Real-time PCR showed that NGF did not influence gene expression of type I, II and XI collagen, TGF-ß, IGF-1 and metalloproteinase (MMP)-13, while it reduced the expression of MMP-3. These findings show that NGF may have uncertain effects in human chondrocytes. Further investigations by wider gene expression and protein synthesis analyses are required to determine how chondrocytes may be influenced by NGF.

Lights and shadows of bone augmentation in severe resorbed mandible in combination with implant dentistry.

Edentulous mandible frequently undergoes severe bone atrophy with problems of prosthetic instability. Instability of the lower denture may cause difficulties with eating and speech, ulcerations of the oral mucosa for lower denture trauma, loss of facial vertical dimension. These problems may be solved by bone augmentation of severe resorption of edentulous mandible. The aim of this short review is to describe surgical techniques for bone augmentation of the severe resorption of edentulous mandible. In this paper, we define a severe resorption of edentulous mandible as a mandibular height in the symphyseal area of 12 mm or less as measured on a standardized lateral cephalogram. Bone grafts and distraction osteogenesis have allowed improving implantology from an experimental to a consolidate dental procedure. It is currently a valuable treatment modality in the prosthetic treatment of severe resorption of edentulous mandible. Numerous techniques have been developed for the rehabilitation of edentulous mandible with fixed or removable mandibular dentures. Today, the options for the restoration of the severe resorption of edentulous mandible with implants can be categorized as follows: insertion of short and narrow implants and a fixed or removable prosthesis; augmentation of the bone with the use of distraction osteogenesis or grafting procedures in combination with the insertion of dental implants loaded with fixed or removable prosthesis; placement of a transosteal dental implants supporting a denture.

Sensory change of the reconstructed breast envelope after skin-sparing mastectomy.

BACKGROUND: Comprehensive studies of somatosensory change following breast reconstruction are limited. We investigated altered sensation quantitatively and qualitatively in patients undergoing mastectomies for cancer treatment and unaffected individuals who had risk-reducing mastectomies (RRM) for cancer predisposing genes. METHODS: Women attending breast clinic review at Royal Marsden Hospital, London were invited to participate. Sensory testing was performed a minimum of 1 year after surgery. Quantitative assessment of light touch and temperature sensation was performed at six points on the breast mound using Semmes-Weinstein monofilaments and temperature regulated droplets. Subjective sensibility of pain, tingling and pleasurable sensation was assessed using a four-point Likert scale questionnaire. RESULTS: 181 breast envelopes were examined, 77 following mastectomy for cancer, 68 after RRM and 36 controls. Partial sensation was maintained with normal light touch in at least 1 quadrant in 57% following surgery. Preserved sensation was highest in the medial breast mound (p = 0.001). On qualitative assessment 74% reported significant loss of pleasurable sensation and 9% reported chronic pain. No difference in light touch and temperature sensation was noted in cancer versus RRM groups but loss of pleasurable sensation was more frequent in the former. Radiotherapy did not affect sensory change post-mastectomy. Following nipple sparing mastectomies, 47% retained normal touch sensation in the preserved areola and nipple. CONCLUSION: Breast sensibility is significantly impaired following mastectomy and reconstruction but sensory loss is partial in the majority of women. Patients should be informed of these adverse post-operative effects to facilitate an informed decision if there is a surgical choice other than mastectomy as a surgical option.

The first case of acinic cell carcinoma of the breast within a fibroadenoma: case report.

A case of acinic cell carcinoma of the breast is reported in a 26-year-old woman. She presented a lump in her right breast, that seemed to be a fibroadenoma. The open biopsy revealed a well-bordered fibroadenoma, together with a proliferation of cells characterized by serous acinar differentiation and eosinophilic cytoplasmic granules. Tumor cells stained for amylase, lysozyme, α-1-antichymotripsin, epithelial membrane antigen, S-100 protein, pan-cytokeratin, cytokeratin 7 and E-cadherin. Estrogen receptor, progesterone receptor, human epidermal growth factor receptor 2 overexpression, CD10, P63, smooth muscle actin, cytokeratin 5/6 were negative. The sentinel node was negative. 8 months after surgery she is in good clinical conditions without recurrence or metastases.

Acinic cell carcinoma of the breast: review of the literature.

INTRODUCTION: The breast and salivary gland tissue share embryologic and thus pathological similarities. Acinic cell carcinoma (ACC) is a typical tumor in salivary glands, but rarely arises in breast too. We reviewed 38 cases of mammary ACC reported in literature and our case, the first ACC born within a fibroadenoma. MATERIALS AND METHODS: Data were collected by a research for the key words acinic cell carcinoma breast on Pubmed in March 2014, including a case treated in our department. All reviewed cases were compared for clinical approach and histological pattern. RESULTS: To date 23 articles presenting cases of ACC of the breast are reported in literature. We included in our review 38 cases previously described and one new case. The histological pattern was predominantly solid with a microglandular structure. All the tumor cells were cytologically characterized by monotonous round cells with a finely granular, weakly eosinophilic, or clearly vacuolated cytoplasm. The most of the cells were intensely stained with anti-lysozime, anti-amylase, anti-α1-chimotripsin, anti-EMA and anti-S100 protein antisera. Immunohistochemistry was also performed to point out: estrogen receptor (ER), progesterone receptor (PR), androgen receptors (AR), human epidermal growth factor receptor 2 overexpression (HER2/neu), E-cadherin (E-cad), cytokeratin-7 (CK7), gross cystic disease fluid protein 15 (GCDFP15), smooth muscle actin (SMA). CONCLUSION: ACC of the breast is a rare tumor, showing similarities with the salivary gland counterpart, above all in terms of good prognosis, and differences from the ordinary invasive breast carcinoma. Further investigations are needed to elucidate the true histogenesis and the correct treatment.

Effects of glucosamine and nucleotide association on fibroblast: extracellular matrix gene expression.

Glucosamine (Gluc) is a drug used as an anti-inflammatory in moderate forms of knee arthrosis. A further off label use of Gluc is in the anti-aging treatments associated with Polideoxirybonucleotide (PDRN) through intra-dermal injection for a procedure called bio-stimulation. An unexpected effect on cultured dermal fibroblasts, during an experimental study on the gene activation in aesthetic bio-stimulation, was observed. The results have potential application in orthopaedic medical therapy. Fibroblast primary cultures were carried out, seeding cells on a layer of Gluc or PDRN alone or in combination for 24 h. Real Time-PCR was performed to investigate several gene expressions. The MMP13 and the IGF-I gene expression in fibroblast cultures were strongly inhibited after 24 h of incubation with the association of Gluc and PDRN, whereas Gluc and PDRN alone produced a modest inhibition of IGF-I and an activation of MMP13. MMP13 is present in osteoarthritic cartilage and this enzyme plays a significant role in cartilage collagen degradation. IGF1 is involved in growth and development and is successfully used in tissue-engineering for cartilage repair. Based on the reported data, we infer that the association of Gluc and PDRN has a potential application in cartilage therapy. Additional basic science and clinical studies are needed to confirm this preliminary report.

Histomorphic-metric evaluation of an implant retrieved from human maxilla after 13 years.

Fixture fracture is the most catastrophic failure of implant components because it usually causes the loss of the implant. Nevertheless, the osseointegrated fractured implants represent a very useful opportunity to study in humans the effects of loading to the peri-implant bone microstructure. The aim of the present study was to evaluate the interplay between microstructure and function of the bone around an implant retrieved from human maxilla after 13 years. There was 1 fractured Dental Implant Line (sand blasted surface from a patient placed in the anterior region of the maxillary bone (2.1) after a bone augmentation procedure, and it was processed for histology. The specimen was analyzed under the scanning electron microscope (SEM), the confocal scanning laser microscope (CSLM) and brightfield light microscope (LM) equipped with circularly polarized light (CPL). The BIC rate of the implant retrieved after 13 years was (mean ±SD) 68.7 ± 3.7. The crestal bone down the implant platform damage appeared to be under modeling process. The transverse collagen fiber orientation (CFO) (mean ±SD) under the lower flank of the threads was 20.4 ± 3.5 x 10(4) pixel while the longitudinal CFO was 19.8 ± 2.8 x 10(4) pixel (P>.05). In the inter-threads region the transverse CFO (mean ±SD) was 15.0 ± 4.0 x 10(4) pixel while the longitudinal CFO was 21.4 ± 3.0 x 10(4) pixel (P>.05). The osteocytes numbers (mean ±SD) was 130 ∓ 34. Under SEM with back scattered electrons (BSE) signal the peri-implant bone appears mainly lamellar and highly mature with several osteons organized in the implant inter-threads areas. The fracture of the implant was most probably correlated to a fatigue of the material mainly associated to a damage of the internal coil. Surprisingly, it was noted a lack of implant site-specific CFO of the bone extracellular matrix facing the threaded dental implant notwithstanding the high level of BIC rate.

Histomorphometric evaluation of an immediately loaded implant retrieved from human mandible after 2 years.

The aim of the present study was to evaluate the interplay between microstructure and function of the bone around an immediately loaded implant retrieved from human maxilla after 23 months due to fracture. A spiral implant of 3.3 mm x 15 mm was placed in a male 53 years old in the anterior region of the mandible bone (4.1) and it was processed for histology. The specimen was analyzed under the confocal scanning laser microscope (CSLM) and brightfield light microscope (LM) equipped with circularly polarized light (CPL). The BIC rate was 76.7 ± 4.9 (mean ±SD). Many cement lines indicates an high remodeling rate of the bone. The transverse collagen fiber orientation (CFO) (mean±SD) under the lower flank of the thread near the tread tip was 55.2 ± 4.8 x 10(4) pixel while the longitudinal CFO was 45.8 ± 2.3 x 10(4) pixel (P<.05). In the inter-threads region the transverse CFO (mean ±SD) was 36.4 ± 2.4 x 10(4) pixel while the longitudinal CFO was 65.6 ± 6.5 x 10(4) pixel (P<.05). The osteocytes numbers (mean ±SD) was 205 ± 45 in the peri-implant bone and 144 ± 53 in the native bone (P=.007). After 2-years of loading the SLA spiral implant was well osseointegrated but still surrounded by woven bone. The osteocytes density was significantly higher in the peri-implant bone than in the native bone. The transverse collagen fibers were significantly associated with the lower flank of the implant threads, while the longitudinal collagen fibers were more represented in the straight surface of the implant. The implant fracture was correlated to crestal bone resorbing and subsequent fatigue yielding.

Calcium sulfate stimulates pulp stem cells towards osteoblasts differentiation.

Calcium sulfate (CaS) is a highly biocompatible material and enhances bone formation in vivo. However, how CaS alters osteoblast activity to promote bone formation is poorly understood. To study how CaS can induce osteoblast differentiation in mesenchymal stem cells, the expression levels of bone related genes and mesenchymal stem cells marker were compared in normal osteoblasts and dental pulp stem cells, using real time Reverse Transcription-Polymerase Chain Reaction. Gene differentially expressed between the two cells type were the trascriptional factor RUNX2, osteopontin (SPP1), COL1A1 (collagen type 1α1) and alkaline phosphatase (ALPL). The obtained results demonstrated that CaS strongly influences the behavior of DPSCs in vitro enhancing proliferation, differentiation and deposition of matrix.

Expansion of the alveolar bone crest with ultrasonic surgery device: clinical study in mandible.

The purpose of this paper was to document the application to the split-crest mandibular procedure in two stage in order to avoid cortical resorption due to periosteal detachment in buccal cortical bone of the alveolar crest. Twenty-two healthy patients with non-contributory past medical history (14 women and 8 men, all non-smokers, mean age 59 years, range 54-65 years) were included in this study. After buccal mucoperiosteal flap was followed by a sagittal corticotomy in the coronal area of the alveolar crest and a second sagittal corticotomy, but in a lower (basal) position and two vertical corticotomies in the buccal wall, using a ultrasonic surgery device (Surgysonic, Esacrom, Imola Italy). Adequate crest expansion was achieved without compromising cortical vascularisation by utilising a combination of scalpel, thin chisels and threaded osteotomes (Bone System, Milano, Italy). Postoperative results were assessed by panoramic and periapical radiographs. Ossification of the osteotomy lines was evident and could be observed as sites with increasing radiopacity on panoramic and periapical radiographs 3 months after implants insertion. No dehiscence of the mucosa was observed. No patient suffered from hypoaesthesia. The mean horizontal bone increase in coronal area was 5±3 mm. Mandibular ridge expansion using a split-crest technique that included grafting the implant sites with a ultrasonic surgery device is a viable therapeutic alternative for implant placement in this patient population.

Assessment of pain associated with insertion torque of dental implants. A prospective, randomized-controlled study.

This study investigated pain experience following dental implant placement in relation to insertion torque using questionnaires. A total 80 implants were placed in 20 patients. Each patient received 4 implants at different times. One implant was inserted and, then, after 40 days was placed the second implant, after 80 days was placed the third implant and after 120 days was placed the fourth implant. At each time the peri-implant bone levels were evaluated on intraoral radiographs taken with the paralleling technique. The implants were placed with a dynamometric key at 35N, 50N, 65N, 85N. Patients were asked to evaluate their pain experience during surgery, 24 hours after surgery, and at 2 days, 4 days, 1, 2, and 4 weeks after surgery on special pain assessment forms. A separate form was used for each time point. Pain was assessed using a descriptive numerical rating scale of 0 to 10, with 0 indicative of no pain and 10 representing the worst pain imaginable. Patients were instructed that a score of 1 to 3 was indicative of mild pain, 4 to 6 was indicative of moderate pain, and 7 to 10 was indicative of severe pain. A significant correlation pain scores and insertion torque was found between group III and group IV vs group II and group I during surgery, at 24 hours, 2 days, 4 days, 1 week, 2 weeks p ≤ 0.05. No statistical difference was found between group I vs. group II during surgery, at 24 hours, 2 days, 4 days, 1 week, 2 weeks p ≥ 0.05. In conclusion, elevated insertion torque values produces pain and resoption of the crestal bone around the implants.

Analysis and structural examination of screw loosening in oral implants.

Biological and technical failures of implants have already been reported. Mechanical factors are certainly of importance in implant failures, even if their exact nature has not yet been established. The abutment screw fracture or loosening represents a rare, but quite unpleasant failure. The aim of the present research is an analysis and structural examination of screw thread or abutment. In this study broken screws were excluded. A total of 58 screw thread loosening were observed, 5 Branemark, (Nobel Biocare, Gothenburg, Sweden) 4 Implant Innovation (Riverside Drive Palm Beach Gardens, FL, USA) and 7 Restore (Lifecore Biomedical, Chaska, Minnesota, USA) and 42 T.B.R. implant (Benax, Ancona, Italy). The loosened abutment screws were retrieved and analyzed under SEM. Many alterations and deformations were present in concavities and convexities of screw threads.

Polylactide-polyglycolide resorbable plates stimulates adipose tissue-derived stem cells towards osteoblasts differentiation.

Polylactide, polyglycolide materials or devices have been utilized routinely during maxillofacial, craniofacial, and orthopaedic reconstructive surgical procedures.(1) These materials combine the benefits of rigid fixation with the advantages of biodegradation, avoiding the need for implant removal and minimizing the risk of other complications.(2) To study how polylactide, polyglycolide acids plates (PLPG plates) can induce osteoblast differentiation and proliferation in mesenchymal stem cells, the expression levels of bone related genes (RUNX2, SP7, ALPL, SPP1, COL1A1, COL3A1 and FOSL1) and mesenchymal stem cells marker (ENG) were measured in adipose derived stem cells (ADSCs) and normal osteoblast (NO) cultivated on PLPG plates after 15 and 30 days of treatment using real time Reverse Transcription-Polymerase Chain Reaction. Significantly differentially expressed genes among ADSCs and NO were SP7, ENG, FOSL1, RUNX, ALPL and SPP1 in the first 15 days of treatment and SP7, ENG FOSL1, COL3A1 COL1A1, SPP1 and ALPL after 30 days. The present study demonstrated that PLPG plates strongly influences the behavior of ADSCs in vitro by enhancing proliferation, differentiation and deposition of matrix.

Total hip arthroplasty using femoral head shelf autografts in dysplastic hips. Our experience.

The present work deals with a series of 28 patients (a total of 31 hips) suffering from coxarthrosis in dysplasia of the hip. All patients had undergone arthroplasty where an autograft of cortico-cancellous bone was taken from the head of the patientís femur. In 21 of these hips the prosthetic components were not cemented while the other 10 cases were. The patients (average age 57 years 5 months) underwent radiography and were clinically evaluated using the Harris Hip Score. The average follow-up was 6 years and 7 months. In the 10 hips which had been cemented, the average pre-operative Harris Hip Score was 46.9 points (range 24-85) while it was 80.5 upon check-up (range 50-98). In the 21 hips which were not cemented the average pre-operative Harris Hip Score was 44.3 points (range 21-62) while, at the time of check-up it was 90.8 (range 60-100). Only in 3 cases (9.67%) did the cotyloid portion require reoperation because of mobilization. In all cases (100%) the graft integrated perfectly with the host bone. The present study provides encouraging data regarding the use of autografts of cortico-cancellous bone taken from the head of the femur during hip replacement in patients with coxarthrosis in dysplasia of the hip. No differences were seen between the cemented and non cemented prostheses.

Modulation of gene expression in human osteoblasts by targeting a distal promoter region of human estrogen receptor-alpha gene.

Estrogen receptor (ER) alpha is expressed during osteoblast differentiation; however, both its functional role in bone metabolism and its involvement in osteoporotic pathogenesis caused by estrogen deficiency are not well understood. Loss of ER alpha gene expression could be one of the mechanisms leading to osteoporosis. Therefore, we investigated a possible modulation of ER alpha gene expression in a human osteoblastic cell line and in four primary osteoblast cultures by using a decoy strategy. Double stranded DNA molecules, mimicking a regulatory region of the ER alpha gene promoter (DNA-102) and acting as a 'silencer' in breast cancer cells, were introduced into osteoblasts as 'decoy' cis-elements to bind and functionally inactivate a putative negative transcription factor, and thus to induce ER alpha gene expression. We found that the DNA-102 molecule was able to specifically bind osteoblast nuclear proteins. Before decoy treatment, absence or variable low levels of ER alpha RNAs in the different cultures were detected. When the cells were transfected with the DNA-102 decoy, an increase in expression of ER alpha and osteoblastic markers, such as osteopontin, was observed, indicating a more differentiated osteoblastic phenotype both in the cell line and in primary cultures. These results showed that the DNA-102 sequence competes with endogenous specific negative transcription factors that may be critical for a decrease in or lack of ER alpha gene transcription. Therefore, osteoblastic transfection with the DNA-102 decoy molecule may be considered a tempting model in a putative therapeutic approach for those pathologies, such as osteoporosis, in which the decrease or loss of ER alpha expression plays a critical role in bone function.

Effects of pulsed electromagnetic fields on human articular chondrocyte proliferation.

Low-energy, low-frequency pulsed electromagnetic fields (PEMFs) can induce cell proliferation in several cell culture models. In this work we analysed the proliferative response of human articular chondrocytes, cultured in medium containing 10% FBS, following prolonged exposure to PEMFs (75 Hz, 2.3 mT), currently used in the treatment of some orthopaedic pathologies. In particular, we investigated the dependence of the proliferative effects on the cell density, the availability of growth factors and the exposure lengths. We observed that PEMFs can induce cell proliferation of low density chondrocyte cultures for a long time (6 days), when fresh serum is added again in the culture medium. In the same conditions, in high density cultures, the PEMF-induced increase in cell proliferation was observed only in the first three days of exposure. The data presented in this study show that the availability of growth factors and the environmental constrictions strongly condition the cellular proliferative response to PEMFs.

A two-locus model for non-syndromic congenital dysplasia of the hip (CDH).

Complex segregation analysis was conducted in a sample of 171 pedigrees collected through probands affected by non-syndromic dysplasia of the hip (CDH) treated in Ferrara's CDH Centre in the period 1991-6. The analysis favoured a two-locus model, in which the accepted segregation model at the major locus was compatible with recessive transmission, with a gene frequency of the deleterious allele of around 0.20. For the other locus, among the Mendelian hypotheses tested, the recessive model turned out to be the most parsimonious. When ultrasonographic level was examined as an indication of severity, a significant improvement in the fit of the model was observed, giving clear evidence that information on the severity of CDH is important for a better definition of the hereditary transmission model.

Effects of pulsed electromagnetic fields on human chondrocytes: an in vitro study.

(3)H-thymidine incorporation was studied in cultured human nasal and articular chondrocytes exposed to low-energy, low-frequency pulsed electromagnetic fields (PEMFs) (75 Hz, 2.3 mT). The reverse transcriptase polymerase chain reaction (RT-PCR) analysis shows that human secondary chondrocytes derived from both nasal and articular cartilage express collagen type II mRNA, which is a specific marker of the chondrocyte phenotype. In a preliminary series of experiments, cells were exposed to PEMF for different time periods ranging from 6 to 30 hours (time-course), in medium supplemented with 10% or 0.5% fetal calf serum (FCS) and in serum-free medium. The ratios between the (3)H-thymidine incorporation in PEMFs and control cultures show an increase of the cell proliferation in cultures exposed to PEMFs when serum is present in the culture medium, whereas no effect was observed in serum-free conditions. The increase in DNA synthesis, induced by PEMFs, was then evaluated only at the times of maximum induction and the results were analyzed by the three-factor analysis of variance (ANOVA). The data presented in this study show that even if (3)H-thymidine incorporation is higher in nasal than in articular chondrocytes, PEMF induce an increase in the proliferation of both cell types. Moreover, the concentration of FCS in the culture medium greatly influences the proliferative response of human chondrocytes to the PEMF exposure. Though normal human osteoblast cells increase their proliferation when exposed to PEMFs if only 10% FCS is present in the medium, human chondrocytes are able to increase their cell proliferation when exposed to PEMFs in the presence of both 0.5% and 10% of FCS in the medium. The results obtained may help to explain the basic mechanisms of PEMF stimulation of fracture healing.

Correlation between pulsed electromagnetic fields exposure time and cell proliferation increase in human osteosarcoma cell lines and human normal osteoblast cells in vitro.

We have exposed cultured bone cells to a pulsed electromagnetic field (PEMF) for different times to find the minimal exposure time necessary to stimulate an increase of DNA synthesis. We used two different human osteosarcoma cell lines, TE-85 and MG-63, and human normal osteoblast cell (NHOC) obtained from surgical bone specimens. The cells were placed in multiwell plates and set in a tissue culture incubator between a pair of Helmoltz coils powered by a pulse generator (1.3-ms pulse, repeated at 75 Hz) for different periods of time. [3H]Thymidine incorporation was used to evaluate cell proliferation. The two osteosarcoma cell lines increase their thymidine incorporation when exposed to a PEMF for at least 30 min, both in a medium containing 10% fetal calf serum and in a serum-free medium. NHOC are known to increase their cell proliferation when exposed to PEMF but only if cultured in the presence of 10% fetal calf serum. In this experimental condition, three of the four cell lineages studied required at least 9 h of PEMF exposure to increase their DNA synthesis, whereas one cell lineage increased its cell proliferation after 6 h of PEMF exposure. Our observations confirm the hypothesis that the proliferative responses of NHOC and human osteosarcoma cell lines to PEMF exposure are quite different. Moreover, NHOC required minimal exposure times to PEMF to increase their cell proliferation, similar to that needed to stimulate bone formation in vivo.

Total hip arthroplasty in octogenarians: our experience.

The progressive aging of the population, a phenomenon that is common to all Western societies, means an increase in the number of patients aged over 80 years requiring total hip replacement. It is the purpose of this study to report our experience in the use of total hip replacement for the treatment of patients aged over 80 years. The study is retrospective, and it was conducted on a group of 27 patients, for a total of 33 hips. An analysis of the results led us to conclude that there are no differences in indications for total hip replacement in patients aged over 80 years as compared to younger subjects. Nonetheless, it is important to carefully evaluate the general conditions of these patients, prior to surgery, because failure may have dramatic consequences.