Background and objectives: Cleft lip palate takes the second place among all anomalies. The complex appearance of cytokines and proliferation markers has still not been clarified despite their possible crucial role in cleft tissue. Therefore, the aim of work was the detection of appearance of pro- and anti-inflammatory cytokines and proliferation marker Ki67, and their inter-correlations in cleft affected lip (CAL). Materials and Methods: The lip material was obtained from 16 children aged before primary dentition during plastic surgery. Control was obtained from 7 non-CAL oral tissue. Tissues were stained for IL-1, IL-4, IL-6, IL-8, IL-10 and Ki67 immunohistochemically. Non-parametric statistic, Mann–Whitney and Spearman’s coefficient were used. Results: All cytokines positive cells were observed more into the epithelium. Statistically significant difference was seen between epithelial IL-1, IL-10, IL-8 and Ki67 positive cells and IL-10-, IL-4-containing connective tissue cells in comparison to the control. Strong positive correlation was detected in CAL epithelium between IL-10 and IL-8, IL-10 and IL-4, IL-10 and IL-1, IL-1 and IL-8, IL-1 and IL-4, IL-4 and IL-8, IL-8 and Ki67, IL-10 and Ki67, but moderate—in connective tissue between IL-1 and IL-10, IL-1 and IL-4. Conclusions: The CAL epithelium is the main source for the interleukins. Rich similar expression of IL-1 and IL-10 suggests the balance between pro-and anti-inflammatory tissue response on basis of dysregulated tissue homeostasis (increase of IL-8). The correlations between the different ILs-1, -4, -8, -10 in CAL epithelium seem to indicate the self-protection compensatory mechanism for intensification of local inflammatory-immune response without involvement of IL-6. The correlations between Ki67 and cytokines indicate the involvement of IL-8 and IL-10 in stimulation of cellular proliferation. IL-4 and IL-10 expression from CAL connective tissue simultaneously to IL-1, IL-4 and IL-10 inter-correlations there suggests the intensification of local immune response regulated probably by main pro-inflammatory cytokine—IL-1.

Background: Strontium (Sr) enriched biomaterials have been used to improve bone regeneration in vivo. However, most studies provide only two experimental groups. The aim of our study was to compare eleven different bone sample groups from osteoporotic and healthy rabbits’ femoral neck, as it is the most frequent osteoporotic fracture in humans. Methods: Osteoporotic bone defects were filled with hydroxyapatite 30% (HA) and tricalcium phosphate 70% (TCP), 5% Sr-enriched HA₃₀/TCP₇₀, HA₇₀/TCP₃₀, or Sr-HA₇₀/TCP₃₀ granules and were compared with intact leg, sham surgery and healthy non-operated bone. Expression of osteoprotegerin (OPG), nuclear factor kappa beta 105 (NFkB-105), osteocalcin (OC), bone morphogenetic protein 2/4 (BMP-2/4), collagen I (Col-1α), matrix metalloproteinase 2 (MMP-2), tissue inhibitor of matrix metalloproteinase 2 (TIMP-2), interleukin 1 (IL-1) and interleukin 10 (IL-10) was analyzed by histomorphometry and immunohistochemistry. Results: Our study showed that Sr-HA₇₀/TCP₃₀ induced higher expression of all above-mentioned factors compared to intact leg and even higher expression of OC, MMP-2 and NFkB-105 compared to Sr-HA₃₀/TCP₇₀. HA₇₀/TCP₃₀ induced higher level of NFkB-105 and IL-1 compared to HA₃₀/TCP₇₀. Conclusion: Sr-enriched biomaterials improved bone regeneration at molecular level in severe osteoporosis and induced activity of the factors was higher than after pure ceramic, sham or even healthy rabbits.

Psoriasis vulgaris is an inflammatory skin condition characterized by dramatic biochemical and immunological changes.The aim of the study was to evaluate antimicrobial response, tissue degeneration reactions and distribution of inflammatory cytokines in untreated psoriatic skin as well as the correlations between these factors and influence on the course of the disease.We evaluated skin samples obtained from routine punch biopsies in 40 patients with psoriasis vulgaris. All tissue specimens were examined by hematoxylin and eosin staining and immunohistochemistry for human beta defensin 2 (HBD-2), matrix metalloproteinase 2 (MMP-2), tumor necrosis factor-alpha (TNF-alpha), interleukin 6 (IL-6) and IL-8. The staining intensity was semi-quantitatively graded.Numerous keratinocytes, fibroblasts and macrophages expressed HBD-2 while the number of MMP-2-positive macrophages, fibroblasts and epitheliocytes varied. TNF-alpha-positive cells varied from a few to numerous in each microscopic field. IL-6-positive cells varied from a few to abundant and IL-8-positive cells from numerous to abundant in each field.This study had a rather small patient number.Psoriatic skin shows a strong correlative increase in skin antimicrobial proteins and enzymes mediating tissue degeneration suggesting that the skin maintains compensatory mechanisms during persistent remodeling. While individual notable decrease in antimicrobial proteins was observed in some tissue samples, generally the increased human beta defensin associated with psoriasis is likely to be due to an altered immune status. TNF-alpha, IL-6 and IL-8 are common cytokines expressed in psoriatic skin plaques to maintain the inflammatory cycle. HBD-2, MMP-2 and TNF-alpha positively correlate with the severity of psoriasis. Meanwhile, the expression of IL-8 significantly decreases with clinically more severe psoriasis, perhaps making these factors candidate prognostic factors for psoriatic inflammation.

The umbilical cord is seen as the main junction between the developing embryo or fetus and placenta. We studied the antimicrobial response, the presence of undifferentiated cells and TGF-α, as well as tissue degeneration and compensatory remodelling processes in the cells of human umbilical cord. Seven umbilical cord tissue samples obtained during premature and full term births were stained with hematoxylin and eosin and by immunohistochemistry for human beta defensin 2 (hBD-2), hematopoietic progenitor cell antigen CD34, matrix metalloproteinase 2 (MMP-2), the tissue inhibitor of matrix metalloproteinase 2 (TIMP-2), nestin and transforming growth factor alpha (TGF-α). The intensity of staining was graded semiquantitatively. Antimicrobial response was more prominent in the Wharton’s jelly – we found numerous hBD-2-containing cells, while hBD-2 positive cells in the walls of arteries and vein varied from moderate to abundance. Numerous cells in the Wharton’s jelly contained CD34, while in the walls of blood vessels few to moderate stained positive for CD34. MMP-2, TIMP-2 and nestin positive cells were found in all the tissue samples and varied from numerous in the walls of blood vessels to abundance in the Wharton’s jelly and the inflammation region. The abundance of TGF-α-containing cells was found in Wharton’s jelly and moderate to numerous cells of blood vessels contained TGF-α. Conclusions: The human umbilical cord possesses antimicrobial activity and shows the presence of undifferentiated cells. The striking distribution and the expression of MMP-2, TIMP-2 and nestin suggest their role in the very effective and extensive tissue degeneration and compensatory remodelling processes. TGF-α seems to be an important growth factor for extra-embryonic tissue development.

Variety of different bone substitutive materials are synthetized to improve bone healing potentials in pathological bone conditions. Physiologically active molecules within biomaterials, can initiate expression level of biomarkers, regulating bone remodeling. Aim of our study was to analyze bone healing process in bone defects followed by implantation with 5% strontium substituted hydroxyapatite (HAP) /tricalcium phosphate (TCP) 70/30 granules (group A) or HAP/TCP biphasic ceramic granules without strontium substitution (group B), or sham surgery affected bone (group C) in osteoporotic rabbits’ femur. Tissue samples from contralateral intact left leg were used for evaluation of systemic effects after surgery. Changes of bone volume were measured and appearance of OPG, NFkB-105, OC, COL-1, BMP-2/4, MMP-2, TIMP-2, IL-1 and IL-10-positive osteocytes in osteoporotic rabbits’ bone defect were evaluated. No statistical difference between groups of trabecular bone volume was detected. All analyzed markers showed higher appearance of positive osteocytes in groups A and B with comparison to control left leg (p<0.05). Only NFkB105-positive cells showed important difference between sham surgery affected leg and control one (p=0.034). Numerous OPG-positive cells appeared in group A, while moderate number of them was found in groups B and C (p=0.025; p=0.027). Numerous to abundant OC-positive osteocytes were detected in group A, while moderate in group C (p=0.034). Statistical difference of rest biomarkers between groups was not detected. We concluded that implantation of biomaterials in osteoporotic bone improves local bone regenerative properties. However, the notable increase of OPG-containing cells proves the increase of osteoclastogenesis suppression and gives the evidence for renew of bone functionality.