Bone marrow cells and CD117-positive haematopoietic stem cells promote corneal wound healing.

PURPOSE: The present study was conducted to evaluate the therapeutic effects of topically applied bone marrow (BM) cells and CD117-positive haematopoietic stem (CD117(+)) cells on alkali-induced corneal ulcers. METHODS: Bone marrow cells and CD117(+) cells were isolated from syngenic mice and labelled with an intracellular cell tracer. Defined corneal wounds were produced in 89 eyes of syngenic mice and allowed to partially heal in vivo for 6 hr. The alkali-burned eyes were enucleated 6 hr postinjury and randomly divided into three groups. Control group (33 eyes) was incubated with medium only. The treatment groups received either BM cells (30 eyes) or CD117(+) cells (26 eyes) suspended in medium. Re-epithelialization process of corneal defects was qualitatively and quantitatively assessed and statistically analysed. The corneas were examined by histological and immunohistochemical methods. RESULTS: We found that the re-epithelialization of corneal wounds in both treatment groups was significantly accelerated as compared to the control group. During the follow-up period (85 hr), the corneal transparency was comparable in all groups. Morphological investigations of corneas from control and treatment group showed no evident differences in the phenotype of the regenerated epithelium. Additionally, corneas in the treatment groups were devoid of donor-derived BM cells and CD117(+) cells, respectively. CONCLUSIONS: This study provides evidence that topical application of BM cells or CD117(+) cells can be used to reconstruct corneal surfaces. Because neither BM cells nor CD117(+) cells were integrated into the corneal epithelium, we suggest that soluble factors could be responsible for the positive effect of BM cells and CD117(+) cells on corneal wound healing.

TFF peptides and mucins are major components of dacryoliths.

PURPOSE: The study was performed to determine whether trefoil factor peptides (TFF) and/or mucins are components of dacryoliths and to gain further insight into dacryolith composition and formation. METHODS: Twenty dacryoliths found in lacrimal surgery in patients suffering from primary acquired nasolacrimal duct obstruction were analyzed for the presence of TFF peptides (TFF1, 2, 3), mucins (MUC1, 2, 3, 4, 5AC, 5B, 6, 7, 8), defense cells (T- and B lymphocytes, macrophages, neutrophils), and antimicrobial substances (alpha defensins 1-3, secretory phospholipase A(2)) by means of light microscopy, histochemistry, immunohistochemistry, reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR. RESULTS: All dacryoliths except one revealed clear immunoreactivity for all three TFF peptides. The immunohistochemical distribution of mucins was inhomogeneous throughout the different dacryoliths. However, in some dacryoliths all mucins investigated were detected. MUC8 showed reactivity in 14 out of 15 dacryoliths analyzed by immunohistochemistry. Most dacryoliths contained alpha defensins 1-3 as the secretory product of neutrophils. T and B lymphocytes, macrophages and secretory phospholipase A(2) were only present in single dacryoliths. Quantification of TFF peptide expression supported the immunohistochemical finding that all three TFF peptides are augmented in dacryoliths. CONCLUSIONS: Dacryoliths consist partly of secreted mucins comparable with the mucin spectrum of the epithelium of healthy nasolacrimal ducts. Beside TFF1 and TFF3, both of which are produced under healthy circumstances, TFF2 is additionally induced and secreted in cases of dacryolithiasis. All three TFF peptides appear to be augmented in dacryoliths. With regard to their rheologic properties, TFF peptides may play a functional role in dacryolith formation. However, our results raise the question of whether TFF peptides per se influence dacryolith formation or whether their secretion, as in secretion of mucins and alpha defensins 1-3, is merely a secondary phenomenon.

Changes in the lens epithelium with respect to cataractogenesis: light microscopic and Scheimpflug densitometric analysis of the cataractous and the clear lens of diabetics and non-diabetics.

BACKGROUND: We compared the human lens documented, using the Scheimpflug densitometry, with the light microscopic changes in the epithelium of the anterior central lens in patients with age-related cataract and diabetes mellitus type II and verified the findings on the control tissue of the clear eye lens. We wanted to determine the relevance of the lens epithelium in cataract formation in type II diabetics compared to non-diabetics. MATERIALS AND METHODS: One hundred fifty central lens capsules (138 cataract and 12 clear lenses) of type II diabetics (n=77, 45 female, 32 male) and non-diabetics (n=73, 41 female, 32 male) were examined by light microscope, regarding defined histomorphological parameters. Further criteria were duration of diabetes, diabetic retinopathy, cataract (PENTACAM, scheimpflug densitometric definition), protein content in the aqueous humour (laser flare meter 500 KOWA, tyndallometry), different blood parameters and glucose content in the aqueous humour. RESULTS: The mean cell density in the cataractous lens in type II diabetics was 3,951+/-528 cells/mm(2) and in non-diabetics 4,329+/-580 cells/mm(2) (P<0.001); in the clear lens it corresponded to 4,593+/-409 cells/mm(2) (type II diabetics) and 4,894+/-333 cells/mm(2) (non-diabetics, P=0.207). The cell density of the cataractous lens in type II diabetics (P=0.005) and in non-diabetics (P=0.035) is smaller than that of the clear lens. The cell area of the lens epithelium in the cataractous diabetic lens is larger (P<0.001) and the nucleus-plasma ratio is lower (P<0.001) than those of the clear non-diabetic lens. The increase in damage of the lens epithelium correlates with the decrease of cell density (P< 0.001), the increase of nucleus area and volume (P< 0.001), and the decrease of nucleus-plasma ratio (P< 0.001). Risk factors for the decrease of cell density are advanced age (P=0.015), type II diabetes (P=0.01), increase in glucose content in the aqueous humour (P=0.014), increase in blood sugar (P=0.003) and increase in glycosylated haemoglobin (P=0.039). CONCLUSIONS: The lens epithelium is primarily damaged in type II diabetics who develop age-related cataract. This might play an important role in cataract formation.

Effects of interleukin-1beta and dexamethasone on the expression of matrix metalloprotease mRNA by trabecular cells exposed to elevated hydrostatic pressure.

PURPOSE: We investigated the effects of interleukin-1beta (Il-1beta) and dexamethasone (Dex) on the expression of matrix metalloprotease-1, -2, -3 and -14 (membrane type-1 MMP-MT1-MMP) as well as tissue inhibitors of matrix metalloproteases (TIMP-1 and -2) mRNA by trabecular cells exposed not only to normal, but also to elevated levels of hydrostatic pressure. METHODS: Confluent primary cultures of porcine trabecular cells were incubated in a serum-free medium (SFM) as controls, or in SFM containing either 10 ng/ml Il-1beta or 10 nm Dex and exposed to pressures of 15 mmHg or 50 mmHg (corresponding to normal and high intraocular pressure, respectively) in specially designed pressure chambers. After 72 hours, total RNA was extracted from the harvested cells, reverse transcribed and amplified using primers specific to MMP-1, -2, -3 and -14, and TIMP-1 and -2. RESULTS: The most significant changes were detected in the levels of MMP-3 mRNA in control cells (2.4-fold increase), of TIMP-1 and -2 mRNA in cells treated with Il-1beta (2.6-fold increase) and of MMP-3 mRNA in cells treated with Dex (3.5-fold increase) exposed to 50 mmHg pressure. CONCLUSION: Because MMP-3 (stromelysin) mRNA showed the highest upregulation, our findings suggest that trabecular cells preferentially degrade and turn over the proteoglycan components of the extracellular matrix in response to short-term exposure to increased hydrostatic pressure with and without Dex as a homeostatic mechanism.

[Hydrophobic AcrySof lenses or heparin-modified PMMA lenses in combined vitrectomy surgery -- results of a four-year study]. / Hydrophobe Acryllinsen (AcrySof) oder heparinmodifizierte PMMA-Linsen bei kombinierten Vitrektomieeingriffen -- Ergebnisse einer 4-Jahres-Studie.

BACKGROUND: Combining pars plana vitrectomy with phacoemulsification and implantation of an intracapsular IOL provides many advantages and is performed as a routine operation. MATERIALS AND METHODS: Over a period of 3 years we compared foldable acrylic lenses AcrySof (Alcon) to heparin-modified PMMA lenses (Pharmacia) in a prospective study. The last implanted lenses had a follow-up period of at least 1 year. Out of 396 eyes from 329 patients we implanted 182 AcrySof lenses and 214 heparin-modified lenses. RESULTS: Both a smaller scleral tunnel incision and a lower rate of secondary cataract are advantages of foldable AcrySof lenses. Higher vulnerability of an AcrySof lens and a less stable lens-iris-diaphragm are of disadvantage. Heparin-modified PMMA lenses do not show these disadvantages, however, in consequence of using them you have to accept the wider scleral tunnel incision and a higher adhesion to cells. Furthermore, we found slightly more posterior synechiae with these lenses. CONCLUSIONS: In all, both types of intraocular lenses are suitable for combined surgery. We recommend using heparin-modified PMMA lenses in case of pre-existent instability of the lens-iris diaphragm or of patients' inability to maintain the prone position.