Differing energy densities with laser 670nm InGaP controls inflammation and collagen reorganization in burns.

PURPOSE: This study compared different energy densities of laser on second degrees burns in rats aiming to determine the most effective dosimetry in stimulation of the healing process. METHODS: Burns were induced in the dorsal skin of 54 animals divided into three groups (n: 18): 1-without treatment; 2-irradiated lesions by the Indium Gallium Phosphide (InGaP) 670nm (4.93J/cm2) laser; 3-irradiated lesions by the InGaP-670nm (9.86J/cm2) laser. Samples were collected on the 2, 10 and 18 days after injury for structural, morphometry, biochemical analysis and Western blotting. RESULTS: The energy densities examined were effective in significantly increasing the total number of fibroblasts and blood vessels and reduce the number of inflammatory cells particularly in irradiated lesions with 9.86J/cm2. This same energy density significantly increased the amount of GAGs (Glycosaminoglycans), decreased the TGF-ß1 (Transforming Growth Factor ß1) and increased the VEGF (Vascular and Endothelial Growth Factor) during the experimental period. This energy density also significantly increased the Collagen type I and decreased Collagen type III and the active isoform of metalloproteinase 9 (MMP-9). CONCLUSIONS: The energy density of 9.86J/cm2 was more effective in promoting cellular responses related to neoangiogenesis, decreasing inflammation and collagen fibers reorganization.

UV irradiation-induced production of monoglycosylated biglycan through downregulation of xylosyltransferase 1 in cultured human dermal fibroblasts.

BACKGROUND: Biglycan (BGN) is a proteoglycan composed of a 42-kDa core protein and two glycosaminoglycan (GAG) chains, and known to be involved in structural, space-filling functions and many physiological regulations in the skin. OBJECTIVE: To investigate ultraviolet (UV) irradiation-induced changes of BGN protein and its GAG chain synthesis in cultured human dermal fibroblasts. METHODS: UV irradiation-induced or xylosyltransferase (XYLT) 1 siRNA-mediated smaller-sized protein bands detected by Western blot using BGN antibodies were identified as monoglycosylated forms of BGN, using BGN siRNA-mediated knockdown and chondroitinase ABC (ChABC). Differential activity of XYLT1 and 2 on BGN core protein was investigated by size shift of S42A- and S47A-BGN mutants to core protein size caused by XYLT1 siRNA transfection or UV irradiation. RESULTS: After UV irradiation, intact form of BGN protein (I-BGN) and core protein form were reduced in cultured fibroblasts, but other smaller-sized bands were observed to be increased. These smaller-sized ones were reduced by transfection of BGN siRNA, and shifted to the core protein size by treatment with ChABC, suggesting that they are defectively-glycosylated forms of BGN (D-BGN) protein. UV irradiation also decreased mRNA expression levels of XYLT1 and 2, which are responsible for initiation of GAG chain synthesis. UV-mediated reduction of XYLT1 expression was much stronger than that of XYLT2. Furthermore, siRNA-mediated down-regulation of XYLT1 resulted in the increase of D-BGN and the decrease of I-BGN, while down-regulation of XYLT2 resulted in no change of D-BGN and I-BGN, suggesting that the XYLT1 may react with both GAG-attaching serine sites of BGN; however, XYLT2 may prefer to react one of them. Another dermatan sulfate (DS) proteoglycan, decorin, showed no or a little change of its molecular weight by UV irradiation or XYLT1 siRNA transfection, suggesting that DS synthesis may not be a critical factor in formation of D-BGN. Co-transfection with XYLT1, 2 siRNAs and wild-type or mutant forms of BGN overexpression vectors revealed that S42A-BGN showed size reduction to core protein size by XYLT1 downregulation, but S47A-BGN did not, suggesting that XYLT2 can react only with S42 on BGN core protein. With UV irradiation, both S42A-BGN and S47A-BGN showed size reduction, which is probably because UV-caused downregulation of both XYLTs and overexpression condition resulted in incomplete glycosylation and secretion. CONCLUSIONS: UV irradiation-induced increase of BGN monoglycosylated forms in cultured human dermal fibroblasts is resulted from dominance of XYLT2 activity, which acts only at S42 on BGN core protein, caused by UV-mediated stronger reduction of XYLT1.

Correlations between X-ray attenuation and GAG content of different cartilage layers based on contrast agent enhanced Micro-CT.

OBJECTIVES: To build the quantitative relationships between X-ray attenuation and glycosaminoglycan (GAG) content of different layers of progressive trypsin digested articular cartilage (AC) models based on the contrast agent enhanced Micro-CT. METHODS: Bovine AC samples were treated with 0.5% concentration of trypsin for different degeneration time, immersed in contrast agent and then scanned by Micro-CT to obtain the X-ray attenuation. Combining with histological analysis, the relationships between the X-ray attenuation and GAG optical density of the superficial and middle layer were analyzed. RESULTS: The X-ray mean attenuation increased about 16.48% (p<;0.01) in the superficial layer within the first 10-minute degeneration, 26.99% (p<;0.05) in the middle layer within the first 40-minute degeneration, but merely changed in the deep layer in the entire experiment. Contrary to the change of X-ray mean attenuation, the GAG optical density decreased from 31.98±13.00 to 11.69±4.23 (p<;0.01) in the superficial layer within the first 10-minute degeneration, from 82.94±7.35 to 3.85±3.31 (p<;0.01) in the middle layer within the entire degeneration and didn't slightly decrease from 96.10±2.50 to 91.45±1.90 (p<;0.05) until the last 10-minute degeneration in deep layer. In addition, the changes of the X-ray mean attenuation showed negative linear correlations with the GAG content in the superficial (r = -0.984, p<;0.01) and the middle layer (r = -0.960, p<;0.01), respectively. CONCLUSION: The contrast agent enhanced Micro-CT can elucidate the variation of GAG content in trypsin-induced progressive AC models by X-ray mean attenuation of different cartilage layers.

Optimization of UV cross-linking density for durable and nontoxic collagen GAG dermal substitute.

Artificial dermal constructs, based upon collagen-glycosaminoglycan matrices (CGMs), provide new options in treating skin defects. However, their clinical effectiveness may be limited by cytotoxicity related to residual aldehydes left over from the manufacturing process. Although both chemical and dehydrothermal (DHT) cross-linking are used to produce CGMs, we hypothesize that optimized nonchemical cross-linking, using ultra-violet (UV) and DHT treatment combinations, may limit cytotoxicity without sacrificing mechanical strength. Porous CGMs were physically cross-linked using a combination of DHT and varying intensities of UV light. These were compared to glutaraldehyde cross-linked controls. Human keratinocytes were seeded in each matrix, and cellular proliferation measured using a microculture tetrazolium dye assay. A scoring system (based on the in vitro contraction rate, stiffness, and cellular growth of a small cylindrical specimen) was developed to assess the best overall physical cross-linking method. More cellular growth was observed in the 90-120 min UV cross-linked group than in the glutaraldehyde-treated group (p < 0.05). Stiffness was maximized after 0-30 min of UV cross-linking. On the basis of our scoring system, DHT combined with 45 min of UV cross-linking produced the best overall matrix in terms of cellular growth and physical durability. UV cross-linked collagen-based biomaterials could be a viable alternative for use in biological applications to eliminate glutaraldehyde-associated cytotoxicity.

Effect of low-dose electron radiation on rat skin wound healing.

The aim of this study was to assess the effect of low-dose electron irradiation on morphological features of the wound healing process in rat skin. Surgical wounds were inflicted with a 2.3 x 1.4 cm template on 84 male rats that were assigned to 4 groups: IG1, immediately irradiated; IG2, irradiated 3 days after inflicting the wound; CG1 and CG2, control groups. Rats in IG1 and IG2 groups had their wounds exposed to 1 Gy of 6 MeV electron beam radiation, immediately after surgery and on the third postoperative day, respectively. Qualitative and histophotometric evaluations of tissue repair structures were carried out. Data were analyzed by ANOVA and Tukey's test (alpha = 0.05) and regression analysis. The repair process was delayed since the first sacrifice time in both irradiated groups, but in IG1, wound healing was closer to that of CG1; whereas in IG2, the delay was more pronounced. Based on the histological findings, it is possible to conclude that a low-dose of electron radiation delayed tissue repair in rat skin. The delay was longer in the skin irradiated 3 days after the beginning of tissue repair. However, the low-energy electron irradiation did not prevent wound healing.

Effect of low-dose electron radiation on rat skin wound healing

The aim of this study was to assess the effect of low-dose electron irradiation on morphological features of the wound healing process in rat skin. Surgical wounds were inflicted with a 2.3 x 1.4 cm template on 84 male rats that were assigned to 4 groups: IG1, immediately irradiated; IG2, irradiated 3 days after inflicting the wound; CG1 and CG2, control groups. Rats in IG1 and IG2 groups had their wounds exposed to 1 Gy of 6 MeV electron beam radiation, immediately after surgery and on the third postoperative day, respectively. Qualitative and histophotometric evaluations of tissue repair structures were carried out. Data were analyzed by ANOVA and Tukey's test (alpha = 0.05) and regression analysis. The repair process was delayed since the first sacrifice time in both irradiated groups, but in IG1, wound healing was closer to that of CG1; whereas in IG2, the delay was more pronounced. Based on the histological findings, it is possible to conclude that a low-dose of electron radiation delayed tissue repair in rat skin. The delay was longer in the skin irradiated 3 days after the beginning of tissue repair. However, the low-energy electron irradiation did not prevent wound healing.

O objetivo deste estudo foi avaliar o efeito da irradiação por baixa dose de elétrons sobre características morfológicas da reparação tecidual na pele de rato. Foram realizadas feridas cirúrgicas com um gabarito de 2,3 x 1,4 cm em 84 ratos machos, divididos em 4 grupos: IG1, irradiado imediatamente; IG2, irradiado 3 dias após a produção da ferida; CG1 e CG2, grupos controles. Os ratos dos grupos IG1 e IG2 tiveram suas feridas expostas a 1 Gy de radiação por elétrons com 6 MeV, imediatamente após a cirurgia e ao terceiro dia pós-operatório, respectivamente. Procedeu-se às avaliações qualitativas e histofotométrica das estruturas de reparação tecidual. Os dados foram submetidos a ANOVA e teste Tukey (a = 0,05) e análise de regressão. O processo de reparação apresentou-se atrasado desde o primeiro tempo de sacrifício em ambos os grupos irradiados, mas para IG1, a reparação tecidual foi similar a CG1. Por outro lado, para IG2, o retardo foi mais pronunciado. Com base nas avaliações histológicas, é possível concluir que uma baixa dose de radiação por elétrons retardou a reparação na pele de rato. O atraso foi maior na pele irradiada 3 dias após o início da reparação tecidual. No entanto, esta irradiação por elétrons de baixa energia não impediu a cicatrização.

Mechanical and biochemical effect of monopolar radiofrequency energy on human articular cartilage: an in vitro study.

BACKGROUND: There are growing concerns about thermal chondroplasty using radiofrequency energy to treat partial-thickness cartilage defects. However, most studies emphasize effects on chondrocyte viability, and other factors such as mechanical properties are less studied. HYPOTHESIS: Radiofrequency energy may cause significant effects on articular cartilage other than chondrocyte viability. STUDY DESIGN: Controlled laboratory study. METHODS: Human osteoarthritic cartilage samples were obtained from total knee arthroplasty, and monopolar radiofrequency energy was applied using commercially available equipment. Material properties (compressive stiffness, surface roughness, and thickness) just before and after thermal treatment were determined using ultrasound. A series of biochemical analyses were also performed after explant culture of the samples. RESULTS: The cartilage surface became smoother by radiofrequency energy, whereas cartilage stiffness or thickness was not altered significantly. Collagen fibrils, especially in the superficial layers, were converted to denatured form, whereas proteoglycan contents released in the media as well as retained in the tissue remained unchanged. The concentrations of matrix metalloproteinases (MMP-1 and MMP-2) were reduced remarkably. CONCLUSION: Radiofrequency energy is able to create a smooth cartilage surface and reduce catabolic enzymes at the cost of collagen denaturation and chondrocyte death in the superficial layers. The stiffness of the cartilage is not changed at time zero. CLINICAL RELEVANCE: Further animal as well as clinical studies will be necessary to fully evaluate the long-term effects of radiofrequency energy.

Effects of helium-neon laser on the mucopolysaccharide induction in experimental osteoarthritic cartilage.

OBJECTIVE: To investigate the effects of mucopolysaccharide induction after treatment by low power laser for experimental osteoarthritis (OA). METHODS: Seventy-two rats with three different degrees of papain induced OA over right knee joints were collected for helium-neon (He-Ne) laser treatment. The severity of induced arthritis was measured by 99mTc bone scan and classified into three groups (I-III) by their radioactivity ratios (right to left knee joints). The rats in each group were further divided into study subgroups (Is, IIs, and IIIs) and control subgroups (Ic, IIc, and IIIc) randomly. The arthritic knees in study subgroups received He-Ne laser treatment, and those in controls received sham laser treatment. The changes of arthritic severity after treatment and follow-up 2 months later were measured. The histopathological changes were evaluated through light microscope after disarticulation of sections (H.E. stain), and the changes of mucopolysaccharide density in cartilage matrix were measured by Optimas scanner analyzer after Alcian blue (AB) stain. The densities of mucopolysaccharide induced after treatment in arthritic cartilage were compared and correlated with their histopathological changes. RESULTS: The density of mucopolysaccharide rose at the initial stage of induced arthritis, and decreased progressively in later stages. The densities of mucopolysaccharide in treated rats increased upon complete laser treatment more than those of the controls, which is closely related with the improvement in histopathological findings, but conversely with the changes in arthritic severity. CONCLUSION: He-Ne laser treatment will enhance the biosynthesis of arthritic cartilage, and results in the improvement of arthritic histopathological changes.

Ergocalciferol promotes in vivo differentiation of keratinocytes and reduces photodamage caused by ultraviolet irradiation in hairless mice.

BACKGROUND: Ergocalciferol (VD(2)) is usually administered orally and it is metabolized to produce its biologically active metabolites in the liver and kidney. Active vitamin D is a well-known potent regulator of cell growth and differentiation. PURPOSE: Active vitamin D such as 1,25-dihydroxyvitamin D(3) (1alpha,25(OH)(2)D(3)) prevents photodamage, including wrinkles and morphologic alterations. However, its clinical and cosmetic use is limited because of its potent, associated effect on calcium metabolism. We examined the efficacy of vitamin D analogues with few adverse effects for preventing skin photodamage. METHOD: Topical application of VD(2) to hairless mouse dorsal skin, and exposure to solar-simulating ultraviolet (UV) radiation at a dose of 10.8 J/cm(2) (UVA) were performed for 15 weeks, five times a week on weekdays. At the end of the final irradiation, histological and analytical studies were performed. RESULTS: Topical application of VD(2) significantly prevented wrinkle formation and abnormal accumulation of extracellular matrix components. In addition, VD(2) suppressed excessive secretion of IL-6 induced by UV irradiation in cultured human normal keratinocytes, in a dose-dependent manner. CONCLUSION: VD(2) promoted keratinocytes differentiation in the epidermis and showed diverse physiological effects, the same as the active form of VD(3). The results suggested that the suppression of skin photodamage involved the promotion of keratinocytes differentiation and suppression of IL-6 secretion induced by exposure to UV. Topical application of VD(2) may become an effective means to suppress solar UV-induced human skin damage.

Collagen loss in photoaged human skin is overestimated by histochemistry.

It is well known that photoaged skin is characterized by increases in dermal matrix components that include glycosaminoglycans, proteoglycans and masses of abnormal elastic fibers accompanied by substantial collagen loss. Histochemical staining of such tissue gives the impression of "massive" loss of collagen and its replacement by these other matrix components. Early biochemical studies have lent support to this notion with a reported decrease in total collagen of approximately 45% compared to protected skin. More recent studies report considerably less, but varying, amounts of collagen loss. Rarely have the two approaches, histochemistry and biochemical analysis, been used in the same study to examine the same tissue. In this study, collagen loss was quantified biochemically in paired biopsies from sun-protected and sun-exposed arm skin of moderately photoaged female subjects (age 51-77 years). The values obtained were compared with histochemical and immunochemical findings. Quantitatively, collagen loss on a per mg protein basis was small compared to the histochemical appearance.

[The effect of laser hemotherapy on the acid glycosaminoglycan content of the liver and blood plasma in rats in the dynamics of an alcoholic coma]. / Vliianie lazernoi gemoterapii na soderzhanie kislykh glikozaminoglikanov v pecheni i plazme krovi krys v dinamike alkogol'noi komy.

The aim of the study was determination of the content of acid glycosaminoglicans (GAG) in hepatic tissue and blood plasma of rats in the course of alcoholic coma and in response to laser irradiation of venous blood. The alcoholic intoxication was achieved by introduction of 40% ethanol through a metal probe into the stomach. The content of acid GAG was defined according to Diche. Alcoholic coma caused reduction of GAG levels in hepatic tissue and an increase in blood plasma which were correlated with the severity of neurological state. Laser irradiation of venous blood promoted a trend to normalization of GAG content in the investigated tissues. A relative reduction in GAG plasma levels after laser hemotherapy indicates effectiveness of the conducted therapy.

The effect of ultraviolet-B exposure scheduling on the photodamage of hairless mouse skin.

In a mouse model, we investigated whether different exposure protocol of ultraviolet-B with the same total doses could induce a different degree of photodamage in mouse skin. Two different exposure frequencies, three times or six times a week, were applied under the condition of weekly same cumulative irradiation dose equally for 10 weeks. Then the photodamage parameters such as skin wrinkling, histochemical dermal change and epidermal and dermal thickness were evaluated. Wrinkle grade, histological assessment by score, and dermal thickness did not reveal any difference between the two groups. However, at irradiation week 10 epidermal thickness of the three times a week irradiation group was significantly thicker than that of the six times a week irradiation group. The same cumulative dose resulted in different epidermal thickness. Our results suggested that exposure frequency or scheduling could influence the epidermal damage by ultraviolet radiation even though the cumulative dose is equal.

Pulsed electromagnetic fields influence hyaline cartilage extracellular matrix composition without affecting molecular structure.

Pulsed electromagnetic fields (PEMF) influence the extracellular matrix metabolism of a diverse range of skeletal tissues. This study focuses upon the effect of PEMF on the composition and molecular structure of cartilage proteoglycans. Sixteen-day-old embryonic chick sterna were explanted to culture and exposed to a PEMF for 3 h/day for 48 h. PEMF treatment did not affect the DNA content of explants but stimulated elevation of glycosaminoglycan content in the explant and conserved the tissue's histological integrity. The glycosaminoglycans in sterna exposed to PEMF were indistinguishable from those in controls in their composition of chondroitin sulfate resulting from chondroitinase ABC digestion. Specific examination with [35S]-sulfate labels showed that PEMF treatment significantly suppressed both the degradation of pre-existing glycosaminoglycans biosynthetically labeled in ovo and the synthesis of new [35S]-sulfated glycosaminoglycans. The average size and aggregating ability of pre-existing and newly synthesized [35S]-sulfated proteoglycans extracted with 4 M guanidinium chloride from PEMF-treated cartilage explants were identical to controls. The chain length and degree of sulfation of [35S]-sulfated glycosaminoglycans also were identical in control and PEMF-treated cultures. PEMF treatment also reduced the amount of both unlabeled glycosaminoglycans and labeled pre-existing and newly synthesized [35S]-sulfated glycosaminoglycans recovered from the nutrient media. [35S]-Sulfated proteoglycans released to the media of both control and PEMF-treated cultures were mostly degradation products although their glycosaminoglycan chain size was unchanged. These results demonstrate that exposure of embryonic chick cartilage explants to PEMF for 3 h/day maintains a balanced proteoglycan composition by down-regulating its turnover without affecting either molecular structure or function.

Photoirritation: a new photobiologic phenomenon induced by long wavelength UVA radiation in hairless mice treated with broad-spectrum sunscreens.

While assessing the protective effect of broad-spectrum sunscreens against chronic UVA radiation, we observed a paradoxical worsening of skin damage with one product. To further examine this finding, five proprietory broad-spectrum sunscreens were applied to albino hairless mice irradiated thrice weekly for 32 weeks with a UVASUN lamp (> 340 nm). Appropriate age-matched controls were included. After approximately 12 weeks, two sunscreens induced a marked dermatitis. Biopsies showed damage greatly exceeding that found in UVA-irradiated, unprotected controls. Histologically, elastic fibers were hyperplastic, coalescing into elastotic clumps. Glycosaminoglycans also increased. Collagen damage was notable since UVA alone does not induce a histologic change. Electron microscopy confirmed these findings. Two other sunscreens provided nearly complete protection. Against chronic UVB radiation, the two UVA photoirritating sunscreens provided substantial protection. Since the UVA sunfilter, oxybenzone, was the same in all sunscreens, we postulate that an irritating component of the vehicle was responsible for the UVA-induced photoirritation. The fifth sunscreen produced severe damage with UVB and UVA.

Effects of radiation on chondrocytes in culture.

Using an in vitro model, we studied the direct effects of radiation on both DNA and glycosaminoglycan (GAG) synthesis of chondrocytes. Chondrocytes from articular cartilage of 21-day-old rabbits were cultured in monolayer or in pellets with matrix as a three-dimensional tube culture. These cells were exposed to a single dose of X-rays at either 2 or 10 Gy. Following irradiation at 2 Gy, DNA synthesis in the chondrocytes was temporarily suppressed but rapidly returned to the control level, while at 10 Gy, DNA synthesis was markedly suppressed and there was no increase in cell number. On the other hand, GAG synthesis was not affected by a single dose of X-rays at either 2 or 10 Gy, nor did GAG content decrease in the three-dimensional tube cultures. These results show that radiation affects the proliferation and differentiation of chondrocytes differently, and that the synthesis of the components of cartilaginous matrix such as GAG is relatively radioresistant in contrast to DNA synthesis.

Alterations of proteoglycans in ultraviolet-irradiated skin.

The effect of UVB exposure on the distribution and synthesis of dermal proteoglycans was measured in the skin of hairless mice. Two groups of mice were included: one was irradiated for 10 weeks; the other was kept as control. After intraperitoneal injection of sodium 35-S-sulfate, punch biopsies were taken for histology and proteoglycans were extracted from the remaining skin with 4 M guanidinium chloride, containing 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (0.5%, weight per volume). Following proteolytic digestion, the glycosaminoglycan constituents were isolated and analyzed by quantitative cellulose acetate electrophoresis and enzymatic digestibility. Under the influence of UVB radiation, newly synthesized proteoglycans measured by 35SO4 uptake increased as much as 60%. In addition, the irradiated skin had a higher average content of proteoglycan than had control skin (4981 micrograms vs 4134 micrograms/g dry weight). This could be ascribed to an increase in heparin (1400 vs 533 micrograms/g dry weight) and heparan sulfate (472 vs 367 micrograms/g dry weight), whereas no change in the concentration of hyaluronic acid (1243 vs 1372 micrograms/g dry weight) and dermatan sulfate (1866 vs 1863 micrograms/g dry weight) was observed. The irradiated animals also exhibited a marked increase in the synthesis of heparan sulfate and heparin (62% and 71%, respectively). These results demonstrate that chronic doses of UVB altered proteoglycan metabolism through both quantitative and qualitative changes.

Dietary vitamin E reverses the effects of ultraviolet light irradiation on rat skin glycosaminoglycans.

Chondroitin 4-sulfate (Ch 4-S), three dermatan sulfates (DS18, DS45a, DS45b) and hyaluronic acid (HA) were the major glycosaminoglycans (GAG) isolated from the skin of 4 groups of albino rats. The yields from Group 1 (control) were: Ch 4-S, 0.015%; HA, 0.028% and DS (total), 0.098% (w/w). Traces of heparin were detected only in rats irradiated with ultraviolet (UV) light (Group II), in the GAG pool isolated with 45% ethanol. Yields increased by at least 28% (w/w) in Group II, but decreased, except HA's, also by at least 28%, below the level of the control, in irradiated rats that also ingested vitamin E (Group III). The sulfate composition of these GAG determined by infrared spectroscopy was as follows: approx. 17% (w/w) for DS18, 21-30% for DS45a, 21-35% for DS45b and 26-44% for Ch 4-S. A 60-70% (mol/mol) N-acetylation of hexosamine in DS45 was estimated by Fourier transform 1H-NMR spectroscopy; the IdUA composition of this DS was 30-46% (mol/mol), and the uronic acid/hexosamine ratio ranged from 2.50:1 to 1.6:1. The data show UV light irradiation of rat skin to result in an abnormally elevated production of the major GAG and oversulfation of Ch 4-S and DS. These effects are reversed, except for the sulfation of DS45b, when the irradiated animals also ingest vitamin E.

Photoinduced prevention of tissue adhesion.

Postoperative tissue adhesion causes retarded wound healing and the need for reoperation; it can even be life threatening. In this report, the authors present prototype materials and performance of newly developed tissue adhesion prevention technology based on photocurable polysaccharides. Polysaccharides used were hyaluronic acid and chondroitin sulfate, both of which were partially derived with photoreactive groups such as cinnamoyl, coumarin, and thymine. Photoreactive hyaluronic acids with low degrees of derivatization were soluble in water. Films of cinnamated hyaluronic acid or an aqueous solution of cinnamated chondroitin sulfate were photocured by ultraviolet irradiation, resulting in water adsorbable films or water swollen gels, respectively. Gelation was due to intermolecular dimerization between cinnamoyl groups. The authors provide two potentially applicable examples: 1) a photocured, water swollen hydrogel film, and 2) a photocurable chondroitin sulfate buffer solution. The authors used hydrogel films to cover the peritoneum after mechanically injuring its surface. Histologic examination showed neither tissue nor cell adhesion, and only a minimal inflammatory response. When tissues were coated with a photocurable chondroitin sulfate solution, the viscous solution was converted to a hydrogel upon ultraviolet irradiation, resulting in in situ tissue covering. Although an optimal molecular design has not yet been found, unique features of mucopolysaccharides (e.g., high water uptake; biodegradability and bioresorbability; and nontoxicity of photodimerizable groups) may result in the development of photoinduced tissue adhesion prevention technology.

Effects of pulsing electromagnetic fields on cultured cartilage cells.

In order to evaluate the effects of pulsing electromagnetic fields (PEMFs) on cell proliferation and glycosaminoglycan (GAG) synthesis and to study the action site of PEMF stimulation in the cells, we performed a series of experiments on rabbit costal growth cartilage cells and human articular cartilage cells in culture. A PEMF stimulator was made using a Helmholz coil. Repetitive pulse burst electric currents with a burst width of 76 ms, a pulse width of 230 microseconds and 6.4 Hz were passed through this coil. The magnetic field strength reached 0.4 mT (tesla) on the average. The syntheses of DNA and GAG were measured by 3H-thymidine and 35S-sulfuric acid incorporations. The effects on the cells treated with lidocaine, adriamycin and irradiation were also measured using a colony forming assay. The PEMF stimulation for the duration of 5 days promoted both cell proliferation and GAG synthesis in growth cartilage cells and intermittent stimulation on and off alternatively every 12 h increased them most significantly, while, in articular cartilage cells, the stimulation promoted cell proliferation, but did not enhance GAG synthesis. PEMF stimulation promoted cells treated with lidocaine more significantly than with other agents. These results present evidence that intermittent PEMF stimulation is more effective on both cell proliferation and GAG synthesis of cartilage cells than continuous stimulation, and that the stimulation could exert effects not by nucleus directly, but by the cellular membrane-dependent mechanism. This study provides further basic data to encourage the clinical application of PEMF stimulation on bone and cartilage disorders.

Wavelength dependence of histological, physical, and visible changes in chronically UV-irradiated hairless mouse skin.

Albino hairless mice (Skh: HR-1) exposed chronically to sub-erythemal doses of UV radiation display physical, visible and histological alterations. Using narrow bandwidth radiation covering the UV radiation spectrum from 280-380 nm, the wavelength dependence of these alterations was determined. The wavelength dependence spectra indicate that for all but one parameter measured (skin sagging), UV-B radiation is considerably more efficient than UV-A radiation in producing changes in the skin. However, in natural sunlight there is considerably more UV-A than UV-B radiation, providing the potential for UV-A to have a larger contribution to skin damage than UV-B. This argues in favor of using broad spectrum photoprotective agents to shield the skin adequately from UV-induced aging. The spectra were also used to develop potential associations among events by determining which events occur at similar wavelengths. There seems to be a correspondence between mouse visible skin wrinking (UV-B event) and two histological events: increase in glycosaminoglycans and alteration in collagen. There was no obvious correspondence among UV-A-induced events.