Regulation of Inflammatory Response in Human Osteoarthritic Chondrocytes by Novel Herbal Small Molecules.

In this study, 34 Traditional Chinese Medicine (TCM) compounds were screened for potential anabolic and anti-inflammatory properties on human osteoarthritic (OA) chondrocytes. The anabolic effects were assessed by measuring the glycosaminoglycan (GAG) relative to the DNA content using a 3D pellet culture model. The most chondrogenic compounds were tested in an inflammatory model consisting of 3 days of treatment with cytokines (IL-1ß/TNF-α) with or without supplementation of TCM compounds. The anti-inflammatory effects were assessed transcriptionally, biochemically and histologically. From the 34 compounds, Vanilic acid (VA), Epimedin A (Epi A) and C (Epi C), 2''-O-rhamnosylicariside II (2-O-rhs II), Icariin, Psoralidin (PS), Protocatechuicaldehyde (PCA), 4-Hydroxybenzoic acid (4-HBA) and 5-Hydroxymethylfurfural (5-HMF) showed the most profound anabolic effects. After induction of inflammation, pro-inflammatory and catabolic genes were upregulated, and GAG/DNA was decreased. VA, Epi C, PS, PCA, 4-HBA and 5-HMF exhibited anti-catabolic and anti-inflammatory effects and prevented the up-regulation of pro-inflammatory markers including metalloproteinases and cyclooxygenase 2. After two weeks of treatment with TCM compounds, the GAG/DNA ratio was restored compared with the negative control group. Immunohistochemistry and Safranin-O staining confirmed superior amounts of cartilaginous matrix in treated pellets. In conclusion, VA, Epi C, PS, PCA, 4-HBA and 5-HMF showed promising anabolic and anti-inflammatory effects.

Improved Adipocyte Viability in Autologous Fat Grafting With Ascorbic Acid-Supplemented Tumescent Solution.

INTRODUCTION: In reconstructive surgery, fat volume augmentation is often necessary for esthetic or functional reasons. As an alternative to synthetic and xenogeneic materials, autologous fat grafting (AFG) based on liposuction is gaining popularity, yet successful transplantation and long-term volume maintenance are difficult. Standard tumescent solution formulations neglect adipocyte and stromal vascular fraction (SVF) cell survival during extraction, as well as SVF differentiation into adipocytes thereafter, all of which are crucial for the success of AFG. Here we hypothesized that addition of ascorbic acid (AA) to the tumescent solution could prevent liposuction-induced cell damage. MATERIALS AND METHODS: The effect of 0.1 mmol/L AA in tumescent solution was investigated in a previously described ex vivo model of AFG. Briefly, excision fat was infiltrated with tumescent solution, with or without AA, and incubated for 20 minutes at 37°C. Hand-assisted liposuction was then performed with a blunt cannula. Total cell viability, clonogenicity, and differentiation capacity of the SVF cells were assessed. RESULTS: With AA, 10.3% more cells and in particular 14.9% more adipocytes survived liposuction. Clonogenicity, adipocyte and osteoblast differentiation by SVF cells remained unchanged. CONCLUSIONS: Addition of AA successfully improved survival of adipocytes during liposuction without affecting SVF growth and differentiation. This study therefore identified a useful supplement to the tumescent solution which may lead to improving AFG success.

Chondrogenic differentiation of human chondrocytes cultured in the absence of ascorbic acid.

Bioreactor systems will likely play a key role in establishing regulatory compliant and cost-effective production systems for manufacturing engineered tissue grafts for clinical applications. However, the automation of bioreactor systems could become considerably more complex and costly due to the requirements for additional storage and liquid handling technologies if unstable supplements are added to the culture medium. Ascorbic acid (AA) is a bioactive supplement that is commonly presumed to be essential for the generation of engineered cartilage tissues. However, AA can be rapidly oxidized and degraded. In this work, we addressed whether human nasal chondrocytes can redifferentiate, undergo chondrogenesis, and generate a cartilaginous extracellular matrix when cultured in the absence of AA. We found that when chondrocytes were cultured in 3D micromass pellets either with or without AA, there were no significant differences in their chondrogenic capacity in terms of gene expression or the amount of glycosaminoglycans. Moreover, 3D pellets cultured without AA contained abundant collagen Types II and I extracellular matrix. Although the amounts of Collagens II and I were significantly lower (34% and 50% lower) than in pellets cultured with AA, collagen fibers had similar thicknesses and distributions for both groups, as shown by scanning electron microscopy imaging. Despite the reduced amounts of collagen, if engineered cartilage grafts can be generated with sufficient properties that meet defined quality criteria without the use of unstable supplements such as AA, bioreactor automation requirements can be greatly simplified, thereby facilitating the development of more compact, user-friendly, and cost-effective bioreactor-based manufacturing systems.

Ascorbic Acid Attenuates Senescence of Human Osteoarthritic Osteoblasts.

The accumulation of senescent cells is implicated in the pathology of several age-related diseases. While the clearance of senescent cells has been suggested as a therapeutic target for patients with osteoarthritis (OA), cellular senescence of bone-resident osteoblasts (OB) remains poorly explored. Since oxidative stress is a well-known inducer of cellular senescence, we here investigated the effect of antioxidant supplementation on the isolation efficiency, expansion, differentiation potential, and transcriptomic profile of OB from osteoarthritic subchondral bone. Bone chips were harvested from sclerotic and non-sclerotic regions of the subchondral bone of human OA joints. The application of 0.1 mM ascorbic acid-2-phosphate (AA) significantly increased the number of outgrowing cells and their proliferation capacity. This enhanced proliferative capacity showed a negative correlation with the amount of senescent cells and was accompanied by decreased expression of reactive oxygen species (ROS) in cultured OB. Expanded cells continued to express differentiated OB markers independently of AA supplementation and demonstrated no changes in their capacity to osteogenically differentiate. Transcriptomic analyses revealed that apoptotic, cell cycle-proliferation, and catabolic pathways were the main pathways affected in the presence of AA during OB expansion. Supplementation with AA can thus help to expand subchondral bone OB in vitro while maintaining their special cellular characteristics. The clearance of such senescent OB could be envisioned as a potential therapeutic target for the treatment of OA.

Maintenance of the response to dimethyl sulfoxide treatment using hyperbaric oxygen in interstitial cystitis/painful bladder syndrome: a prospective, randomized, comparative study.

INTRODUCTION: Interstitial cystitis (IC)/painful bladder syndrome (PBS) is a difficult disease to manage and creates critical limitations in patients' daily lives. Our objective was to determine the efficacy of hyperbaric oxygen (HBO) therapy in the maintenance of response after the administration of intravesical dimethyl sulfoxide (DMSO). MATERIALS AND METHODS: We conducted an open, prospective, randomized, comparative pilot study with women diagnosed with IC/PBS according to the European Society for the Study of Interstitial Cystitis criteria. In the first phase, DMSO was given to all patients. In the second phase, we used 1:1 randomization and administered HBO to 10 women. The evaluated variables were pain (through a visual analog scale), frequency and urgency of voids, nocturia, and quality of life using the O'Leary-Sant Interstitial Cystitis Score/Problem Index and the King's Health Questionnaire. In the second phase, we measured the length of time that clinical improvement was maintained. RESULTS: The mean age was 47.6 years (SD 18.4). Out of 20 patients, 14 experienced clinical improvement after DMSO in all of the evaluated symptoms (p < 0.05; 95% CI). After the second phase, all patients who received HBO had a more substantive and prolonged maintenance of the effects of DMSO. CONCLUSIONS: In this study, HBO improved the maintenance of the beneficial effects of DMSO among women with IC/PBS.

Interleukin-1ß modulates endochondral ossification by human adult bone marrow stromal cells.

Inflammatory cytokines present in the milieu of the fracture site are important modulators of bone healing. Here we investigated the effects of interleukin-1ß (IL-1ß) on the main events of endochondral bone formation by human bone marrow mesenchymal stromal cells (BM-MSC), namely cell proliferation, differentiation and maturation/remodelling of the resulting hypertrophic cartilage. Low doses of IL-1ß (50 pg/mL) enhanced colony-forming units-fibroblastic (CFU-f) and -osteoblastic (CFU-o) number (up to 1.5-fold) and size (1.2-fold) in the absence of further supplements and glycosaminoglycan accumulation (1.4-fold) upon BM-MSC chondrogenic induction. In osteogenically cultured BM-MSC, IL-1ß enhanced calcium deposition (62.2-fold) and BMP-2 mRNA expression by differential activation of NF-κB and ERK signalling. IL-1ß-treatment of BM-MSC generated cartilage resulted in higher production of MMP-13 (14.0-fold) in vitro, mirrored by an increased accumulation of the cryptic cleaved fragment of aggrecan, and more efficient cartilage remodelling/resorption after 5 weeks in vivo (i.e., more TRAP positive cells and bone marrow, less cartilaginous areas), resulting in the formation of mature bone and bone marrow after 12 weeks. In conclusion, IL-1ß finely modulates early and late events of the endochondral bone formation by BM-MSC. Controlling the inflammatory environment could enhance the success of therapeutic approaches for the treatment of fractures by resident MSC and as well as improve the engineering of implantable tissues.

Effects of a perfusion bioreactor activated novel bone substitute in spine fusion in sheep.

PURPOSE: To evaluate the effect of a large perfusion-bioreactor cell-activated bone substitute, on a two-level large posterolateral spine fusion sheep model. METHODS: A 50 mm long porous biphasic-calcium-phosphate bone substitute reinforced with poly(D,L-lactide) and, activated with bone marrow derived mononuclear-cells (BMNC) was used. Eighteen sheep were divided into two groups and one group (n = 9) had BMNC-activated bone substitutes and cell-free substitutes implanted. The second group (n = 9) had autograft supplemented with BMNC and regular autograft implanted. The implant material was alternated between spine level L2-L3 and L4-L5 in both groups. MicroCT was used to compare the spine fusion efficacy and bone structure of the two groups as well as the implanted bone substitutes and non-implanted substitutes. RESULTS: After 4½ months six sheep survived in both groups and we found five spine levels were fused when using activated bone substitute compared to three levels with cell-free bone substitute (p = 0.25). Five sheep fused at both levels in the autograft group. A significant increased bone density (p < 0.05) and anisotropy (p < 0.05) was found in the group of activated bone substitutes compared to cell-free bone substitute and no difference existed on the other parameters. The implanted bone substitutes had a significant higher bone density and trabecular thickness than non-implanted bone substitutes, thus indicating that the PLA reinforced BCP had osteoconductive properties (p < 0.05). No effect of the supplemented BMNC to autograft was observed. The autograft group had a significant higher bone density, trabecular thickness and degree of anisotropy than the implanted bone substitutes (p < 0.05), but a lower connectivity density existed (p < 0.05). This indicates that though the activated substitute might have a similar fusion efficacy to autograft, the fusion bridge is not of equal substance. CONCLUSION: We found that bioreactor-generated cell-based bone substitutes seemed superior in fusion ability when compared to cell-free bone substitute and comparable to autograft in fusion ability, but not in bone structure. This combined with the favorable biocompatible abilities and strength comparable to human cancellous bone indicates that it might be a suitable bone substitute in spine fusion procedures.

In vivo UVB-photoprotective activity of extracts from commercial marine macroalgae.

The photoprotective potential against UVB radiation of extracts obtained from 21 commercial macroalgae from the Phyla Ochrophyta and Rhodophyta, was evaluated in vivo, using the zebrafish embryo as a whole model organism. Our results showed that the phenolic extracts from Macrocystis pyrifera and Porphyra columbina exhibited the highest photoprotective activity, close to complete photoprotection (100%), similar to that obtained for the carrageenophytes Sarcothalia radula and Gigartina skottsbergii. Under the assayed conditions, the extracts were safe and non-toxic to the embryos at a concentration of 0.04 mg/ml PGE.

Hyperbaric oxygen therapy for the management of hemorrhagic radio-induced cystitis.

OBJECTIVES: Radio-induced cystitis (RADC) is an inflammatory bladder disease that presents as anemic-hematuria in its most serious form. Classic treatments can not control the disease in the mid-to-long term because they don't treat the pathogenesis of the disease. Thus, we evaluated the effectiveness of hyperbaric oxygen (HBO) therapy as a potential treatment for patients with RADC. METHODS: This prospective study included 38 patients, 21 men and 17 women, mean age of 66.5 years(46-75), who had been subjected to pelvic radiotherapy (RT), with the diagnosis of RADC with or without radio-induced proctitis (RADP), gross hematuria and lower urinary tract symptoms. HBO treatment was applied in a multiplace chamber; patients breathed pure oxygen (100%) at 2-2.5 atmospheres of pressure (ATAs). Patients received an average of 31.2 sessions (10-48 sessions) and the median follow-up period was 56 months (4-72 months). RESULTS: Hematuria was completely resolved in 34 of the 38 patients. After HBO 6 patients required readmission, 5 for anemic hematuria and 1 for acute obstructive pyelonephritis. In general, patients tolerated treatment well; however, one patient experienced barotrauma requiring myringotomy. CONCLUSIONS: HBO can be used to satisfactorily treat RADC, leading to clinical improvements that begin during the initial sessions in the majority of cases, and with a more than acceptable level of patient tolerance.

Oxigenoterapia Hiperbárica (OHB) en el manejo de la cistitis hemorrágica radioinducida / Hyperbaric Oxygen Therapy for the management of hemorrhagic radio-induced cystitis

OBJETIVO: La Cistitis rádica (CRAD) es una enfermedad inflamatoria vesical que se presenta de forma más grave como hematuria anemizante. Los tratamientos clásicos no consiguen controlar la enfermedad a medio-largo plazo ya que no actúan sobre su patogénesis. Evaluamos la respuesta clínica de pacientes con cistitis radioinducida tras ser tratados mediante Oxigenoterapia Hiperbárica.MÉTODOS: Estudio prospectivo en el que se incluyen 38 pacientes, 21 hombres y 17 mujeres, edades desde los 46 a los 75 (media de edad de 66.5 años) sometidos a radioterapia (RT) pélvica, diagnosticados de CRAD +/- proctitis radioinducida (PRAD) y que clínicamente referían hematuria, y síndrome miccional. El tratamiento se aplicó en una cámara de tipo multiplaza, los pacientes respiraban O2 al 100% a una presión ambiental de 2-2,5 ATAs (atmósferas de presión ambiental). Recibieron una media de 29.9 sesiones (rango 10-48 sesiones), el seguimiento medio fue de 56 meses (rango 4-72 meses).RESULTADOS: La hematuria se ha resuelto hasta la fecha de forma completa en 35 pacientes, un paciente presenta actualmente hematurias no anemizantes ocasionales, a razón de una 1 trimensual. Requiriendo reingreso 6 de ellos, 5 por hematuria anemizante y 1 por pielonefritis aguda obstructiva. El tratamiento fue bien tolerado por los pacientes, 1 experimentó barotrauma que requirió de miringotomía.CONCLUSIONES: La CRAD puede tratarse de forma satisfactoria mediante OHB, consiguiendo mejoría clínica, desde las primeras sesiones en la mayoría de ocasiones, con una tolerancia más que aceptable por parte de los pacientes(AU)

OBJECTIVES: Radio-induced cystitis (RADC) is an inflammatory bladder disease that presents as anemic-hematuria in its most serious form. Classic treatments can not control the disease in the mid-to-long term because they don`t treat the pathogenesis of the disease. Thus, we evaluated the effectiveness of hyperbaric oxygen (HBO) therapy as a potential treatment for patients with RADC.METHODS: This prospective study included 38 patients, 21 men and 17 women, mean age of 66.5 years (46-75), who had been subjected to pelvic radiotherapy (RT), with the diagnosis of RADC with or without radio-induced proctitis (RADP), gross hematuria and lower seurinarytract symptoms. HBO treatment was applied in a multi-place chamber; patients breathed pure oxygen (100%) at 2-2.5 atmospheres of pressure (ATAs). Patients received an average of 31.2 sessions (10-48 sessions) and the median follow-up period was 56 months (4-72 months).RESULTS: Hematuria was completely resolved in 34 of the 38 patients. After HBO 6 patients required readmission, 5 for anemic hematuria and 1 for acute obstructive pyelonephritis. In general, patients tolerated treatment well; however, one patient experienced barotrauma requiring myringotomy.CONCLUSIONS: HBO can be used to satisfactorily treat RADC, leading to clinical improvements that begin during the initial sessions in the majority of cases, and with a more than acceptable level of patient tolerance(AU)

Empleo de la oxigenoterapia hiperbárica en Urología / Hypebaric oxygen treatment in urology

OBJETIVO: La oxigenoterapia hiperbárica (OHB) se ha empleado de forma existosa en numerosas patologías que derivan de la hipoxia tisular gracias al aporte extra de oxígeno que permite a los tejidos.En este trabajo se realiza una revisión exhaustiva acerca de toda la literatura existente en 2010 en la que se emplea OHB en patología urológica.MÉTODOS: Realizamos una búsqueda en Medline introduciendo los términos “hyperbaric oxygen”, “radic cistitis”, “interstitial cistitys”, “ hemorraghic cistitys”, “urological/pelvic fistula” y “Fournier´s gangrene”.Las búsquedas se centraron en estudios en humanos únicamente publicados en cualquier idioma.RESULTADOS: 56 trabajos publicados, 1 ensayo clínico controlado aleatorizado (ECA), 7 revisiones (review) y 48 series de casos (SC) de los que tan solo uno fué prospectivo en los que se exponen a un total de 695 pacientes. Sólo en un estudio se emplearon mediciones de oxígeno tisular para definir la hipoxia. El número de las sesiones de terapia de oxígeno hiperbárico varió desde 4 hasta 44 sesiones. (media 19,2 sesiones/paciente)CONCLUSIONES: La evidencia que se extrae de la mayoría de trabajos consultados procede de series de casos, de modo que es baja, sin embrago, en la mayoría de estudios los resultados en cuanto al manejo de los pacientes es bueno o muy bueno así que parece que la OHB puede ser de gran utilidad en enfermedades urológicas que deriven de hipoxia tisular(AU)

OBJECTIVES: Hyperbaric oxygen therapy (HBO) has been successfully used in several disorders derived from tissue hypoxia, due to the extra oxygen supply to the tissues it enables.In this manuscript we performed a systematic review including all the existing data published until 2010 about HBO in urologic disorders.METHODS: We performed a Medline search using the terms “hyperbaric oxygen”, “radical cystitis”, “interstitial cystitis”, “hemorrhagic cystitis”, “urological/pelvic fistula” and “Fournier´s gangrene”. The search was restricted to human clinical trials published in any language. RESULTS: We found 56 papers: 1 randomized controlled trial, 7 reviews and 48 case reports; only one of them was a prospective study. A total of 695 patients were included. Just one study used tissue oxygen measurement to define hypoxia. The number of hyperbaric oxygen therapy sessions ranged from 4 to 44 (mean 19.2 sessions/patient).CONCLUSIONS: The level of evidence from most reviewed papers is low because most of them are case series. Nevertheless, results of most of those studies regarding patient management are good or very good. So it seems that HBO can be very useful in urological diseases related to tissue hypoxia(AU)

Hypebaric oxygen treatment in urology.

OBJECTIVES: Hyperbaric oxygen therapy (HBO) has been successfully used in several disorders derived from tissue hypoxia, due to the extra oxygen supply to the tissues it enables. In this manuscript we performed a systematic review including all the existing data published until 2010 about HBO in urologic disorders. METHODS: We performed a Medline search using the terms "hyperbaric oxygen", "radical cystitis", "interstitial cystitis", "hemorrhagic cystitis", "urological/pelvic fistula"and "Fournier's gangrene". The search was restricted to human clinical trials published in any language. RESULTS: We found 56 papers: 1 randomized controlled trial, 7 reviews and 48 case reports; only one of them was a prospective study. A total of 695 patients were included. Just one study used tissue oxygen measurement to define hypoxia. The number of hyperbaric oxygen therapy sessions ranged from 4 to 44 (mean 19.2 sessions/patient). CONCLUSIONS: The level of evidence from most reviewed papers is low because most of them are case series. Nevertheless, results of most of those studies regarding patient management are good or very good. So it seems that HBO can be very useful in urological diseases related to tissue hypoxia.

Anabolic and catabolic responses of human articular chondrocytes to varying oxygen percentages.

INTRODUCTION: Oxygen is a critical parameter proposed to modulate the functions of chondrocytes ex-vivo as well as in damaged joints. This article investigates the effect of low (more physiological) oxygen percentage on the biosynthetic and catabolic activity of human articular chondrocytes (HAC) at different phases of in vitro culture. METHODS: HAC expanded in monolayer were cultured in pellets for two weeks (Phase I) or up to an additional two weeks (Phase II). In each Phase, cells were exposed to 19% or 5% oxygen. Resulting tissues and culture media were assessed to determine amounts of produced/released proteoglycans and collagens, metalloproteinases (MMPs), collagen degradation products and collagen fibril organization using biochemical, (immuno)-histochemical, gene expression and scanning electron microscopy analyses. In specific experiments, the hypoxia-inducible factor-1alpha (HIF-1alpha) inhibitor cadmium chloride was supplemented in the culture medium to assess the involvement of this pathway. RESULTS: Independent from the oxygen percentage during expansion, HAC cultured at 5% O(2) (vs 19% O(2)) during Phase I accumulated higher amounts of glycosaminoglycans and type II collagen and expressed reduced levels of MMP-1 and MMP-13 mRNA and protein. Switching to 19% oxygen during Phase II resulted in reduced synthesis of proteoglycan and collagen, increased release of MMPs, accumulation of type II collagen fragments and higher branching of collagen fibrils. In contrast, reducing O(2) during Phase II resulted in increased proteoglycan and type II collagen synthesis and reduced expression and release of MMP-13 mRNA and protein. Supplementation of cadmium chloride during differentiation culture at 5% O(2) drastically reduced the up-regulation of type II collagen and the down-regulation of MMP-1 mRNA. CONCLUSIONS: The application of more physiologic oxygen percentage during specific phases of differentiation culture enhanced the biosynthetic activity and reduced the activity of catabolic enzymes implicated in cartilage breakdown. Modulation of the oxygen percentage during HAC culture may be used to study pathophysiological events occurring in osteoarthritis and to enhance properties of in vitro engineered cartilaginous tissues.

Three-dimensional perfusion culture of human bone marrow cells and generation of osteoinductive grafts.

Three-dimensional (3D) culture systems are critical to investigate cell physiology and to engineer tissue grafts. In this study, we describe a simple yet innovative bioreactor-based approach to seed, expand, and differentiate bone marrow stromal cells (BMSCs) directly in a 3D environment, bypassing the conventional process of monolayer (two-dimensional [2D]) expansion. The system, based on the perfusion of bone marrow-nucleated cells through porous 3D scaffolds, supported the formation of stromal-like tissues, where BMSCs could be cocultured with hematopoietic progenitor cells in proportions dependent on the specific medium supplements. The resulting engineered constructs, when implanted ectopically in nude mice, generated bone tissue more reproducibly, uniformly, and extensively than scaffolds loaded with 2D-expanded BMSCs. The developed system may thus be used as a 3D in vitro model of bone marrow to study interactions between BMSCs and hematopoietic cells as well as to streamline manufacture of osteoinductive grafts in the context of regenerative medicine.

In vitro and in vivo evaluation of differentially demineralized cancellous bone scaffolds combined with human bone marrow stromal cells for tissue engineering.

Mineralized and partially or fully demineralized biomaterials derived from bovine bone matrix were evaluated for their ability to support human bone marrow stromal cell (BMSC) osteogenic differentiation in vitro and bone-forming capacity in vivo in order to assess their potential use in clinical tissue-engineering strategies. BMSCs were either seeded on bone-derived scaffolds and cocultured in direct cell-to-scaffold contact, allowing for the exposure of soluble and insoluble matrix-incorporated factors, or cocultured with the scaffold preparations in a transwell system, exposing them to soluble matrix-incorporated factors alone. Osteoblast-related markers, alkaline phosphatase (ALP) activity and bone sialoprotein (BSP) and osteopontin (OP) mRNA expression were evaluated in BMSCs following 14 days of cocultivation in both systems. The data demonstrate that BMSCs from some donors express significantly higher levels of all osteoblast-related markers following cocultivation in direct cell-to-scaffold contact with mineralized scaffolds in comparison to fully demineralized preparations, while BMSCs from other donors display no significant differences in response to various scaffold preparations. In contrast, BMSCs cocultured independently with soluble matrix-incorporated factors derived from each scaffold preparation displayed significantly lower levels of ALP activity and BSP mRNA expression in comparison to untreated controls, while no significant differences were observed in marker levels between cells cocultured similarly with different biomaterial preparations. In addition, BMSCs were seeded directly on mineralized and partially or fully demineralized biomaterials and implanted in subcutaneous sites of athymic mice for 8 weeks to evaluate their in vivo bone-forming capacity. The ex vivo incorporation of BMSCs into all bone-derived scaffold preparations substantially increased the mean extent and frequency of samples containing de novo bone formation over similar nonseeded controls, as determined by histological and histomorphometrical analysis. No statistically significant differences were observed in the extent or frequency of bone formation between various scaffold preparations seeded with BMSCs from different donors. These results demonstrate that the in vivo osteoinductivity of bone-derived scaffolds can be modulated by ex vivo incorporated BMSCs and the extent of scaffold demineralization plays a significant role in influencing in vitro osteogenic differentiation of BMSCs depending on the coculture system and BMSC donor.

FGF-2 enhances TGF-beta1-induced periosteal chondrogenesis.

The use of periosteum as a cell source for the in vitro engineering of grafts for articular cartilage repair requires the development of methods to obtain high viable cell numbers in the early stages of culture. In this study, we demonstrate that the addition of a mitogen, fibroblast growth factor-2 (FGF-2), during the early stage of the in vitro culture of periosteum in the presence of transforming growth factor-beta1 (TGF-beta1), significantly enhances cell proliferation, which results in increased neo-cartilage formation at later stages. Periosteal explants were cultured in vitro within alginate or agarose based gels in the presence of either FGF-2 for the first week, TGF-beta1 for the first 2 weeks, FGF-2 and TGF-beta1 for the first week and first 2 weeks respectively, or no added factors. Consistent with previous studies, periosteum derived neo-chondrogenesis occurred only in the presence of TGF-beta1. The neo-cartilage was found to contain cartilage specific proteoglycans and Type-II collagen as determined by safranin-O and immunohistochemical staining respectively. Further medium supplementation with FGF-2 stimulated early cell proliferation (>3 fold higher total DNA content per explant at day 10). This resulted in a marked increase in the size of the cultured explants and in the total area of the explant staining positive for safranin-O (from around 50% to 85%, (p<0.05)) after 6 weeks culture. The ability to generate significant quantities of neo-cartilage within a biocompatible and biodegradable matrix such as alginate, which lacks the immunogenicity of agarose, could open new pathways to utilizing such constructs in articular cartilage tissue engineering applications.

Auriculoterapia. Metodología y resultados preliminares

Se realiza un estudio de 125 pacientes tratados con auriculopuntura en el Servicio de Medicina Física y Rehabilitación del Hospital Provincial Docente Vladimir Ilich Lenin, en el período comprendido de mayo a octubre de 1989. Se ofrece un breve resumen de la metodología de aplicación. Se ofrecen resultados preliminares obtenidos