Long-term results of autologous submandibular gland transfer for the surgical treatment of severe keratoconjunctivitis sicca.

AIM: The aim of this study was to assess the long-term results of autologous submandibular gland transfer for surgical correction of severe keratoconjunctivitis sicca. PATIENTS AND METHODS: A survey was undertaken of 32 patients who had undergone submandibular gland transfer (42 glands) and by following up 11 patients (15 glands) for 5-10 years. Subjective benefit was evaluated as well as clinical findings at the ocular surface. The biochemical consistency of the secreted "saliva-tear" was analysed and compared with natural submandibular saliva of a matched control-group. The vitality and function of the transplants was tested by means of sialoscintigraphy. Immunohistochemical investigations were carried out in specimens of submandibular tissue, gained during reduction procedures of the transplants to correct secretory excess. RESULTS: Patient evaluation and clinical assessment revealed a long-lasting subjective benefit in 2/3 of the patients and a stabilisation at the ocular surface in all cases. The secretion remained as highly concentrated submandibular saliva. Glandular vitality and function was shown scintgraphically. Immunohistochemical investigations revealed no progressive atrophy after transplantation, the ability of cell division remained intact and there was still neuronal tissue in all transplants, even several years after transfer. As all transplants responded well to parasympathomimetic drugs, this might be an indication of re-innervation of the gland.

Innervation and secretory function of transplanted human submandibular salivary glands.

Free submandibular gland autotransplantation is used to treat absolute tear deficiency. Although disconnected from any peripheral innervation, most transplants show increasing secretion for years. We have evaluated the secretory activity and autonomic innervation of such transplants. Secretory activity of glands in response to parasympatholytics and parasympathomimetics was evaluated by Schirmer's test and Technetium scintigraphy. Submandibular gland tissue specimens taken before and after transplantation were examined histologically. Relative hypersecretion during the first postoperative week suddenly decreased but then slowly increased during the first postoperative year. Hypersecretion was significantly reduced by parasympatholytics while carbachol rapidly increased secretion. Histology of transplanted glandular tissue showed parenchymal atrophy. Cholinesterase-positive nerves were abundant and in a similar distribution to normal with scattered positive ganglion cells. Adrenergic axons were fewer than normal and irregularly distributed. Early hypersecretion may be due to release of neurotransmitters from degenerating terminal axons. This is followed by a period of minimal secretion during which hypersensitivity of acinar cells develops. With spontaneous reinnervation, secretion is accentuated by external sympathetic vasomotor adrenergic drive. This shows that submandibular glands can remain viable despite complete separation from their normal nerve supply and are capable of regaining a substantial secretory activity for years.

Early and late immunohistochemical and ultrastructural changes associated with functional impairment of the lachrymal gland following external beam radiation.

The aim of this study was to investigate scintigraphic, immunohistological and ultrastructural changes associated with radiation-induced dysfunction of the lachrymal gland in an established experimental animal model. Ten rabbits were randomized into two groups and used for the study; in the control as well as experimental group, the Schirmer-test, lachrymal gland scintigraphy, and immunohistological and ultrastructural investigations were carried out prior to irradiation and 72 h as well as 1 month after single-dose irradiation with 15 Gy. Seventy-two hours after irradiation, secretion reduction evaluated by the Schirmer-test was evident. At this phase, we could observe a decrease in the expression of alpha-SMA and a re-distribution of tenascin-C matrix. Ultrastructural changes of acinar and myoepithelial cells were noticed; simultaneously, disturbance in the primary (99m)TcO(4)- uptake as well as significant reduction of the lachrymal ejection fraction was assessed scintigraphically. These changes were still evident 1 month following irradiation but became less intensive. Single-dose irradiation with 15 Gy implicates a functional impairment of the lachrymal gland, which is associated with early immunohistological and ultrastructural alterations. These changes may represent objective surrogate parameters for radiogenic dysfunction and prerequisites for further investigations on radioprotection of lachrymal glands during radiotherapy of the periorbital region.

A comparative study on the protection profile of lidocaine, amifostine, and pilocarpin on the parotid gland during radiotherapy.

The aim of this study was to evaluate the individual and the synergetic radioprotective effect of lidocaine, amifostine, and pilocarpin on the parotid gland. Forty-nine rabbits were randomized into seven groups (n = 7)--control, irradiated sham-treated, irradiated/lidocaine-pretreated, irradiated/amifostine-pretreated, irradiated/pilocarpin-pretreated, irradiated/lidocaine + pilocarpin-pretreated, and irradiated/amifostine + pilocarpin-pretreated groups. One week before irradiation (15 Gy) and 72 hours as well as 1 month afterward, the parotid gland was investigated morphologically, sialoscintigraphically, and immunohistochemically with the use of tenascin-C and alpha smooth muscle actin. Compared with control animals, there was a significant reduction of the salivary ejection fraction in the irradiated untreated group 72 hours following radiation. Only animals pretreated with lidocaine or amifostine (alone or combined with pilocarpin) showed a slight nonsignificant reduction of salivary ejection fraction. Immunohistochemically, we observed a significant loss of alpha smooth muscle actin and an up-regulation of tenascin-C expression in irradiated/untreated glands. These changes were less evident in animals pretreated with lidocaine or lidocaine + pilocarpin. Amifostine and pilocarpin did not show any influence on tenascin-C or alpha smooth muscle actin expression. Ultrastructural damage was observed in irradiated untreated and pilocarpin-pretreated glands. In contrast, lidocaine and amifostine could largely preserve the glandular ultrastructure. One month postradiation, all changes were regressive regardless of treatment protocol. Potential radioprotective agents show different effects on both morphology and function of the parotid gland. Associated immunohistochemical and ultrastructural findings could prove the prevailed protection profile of lidocaine. This may provide a prophylactic approach in the field of radioprotection of salivary glands.

Monitoring of a new approach of immunotherapy with allogenic (111)In-labelled NK cells in patients with renal cell carcinoma.

The transfusion of allogenic, in vitro expanded natural killer cells (NKC) is a novel therapy option in oncology. To date, however, the biodistribution and kinetics of allogenic NKC have not been investigated. Therefore, in this study three patients with renal cell carcinoma received 3-7 x 10(8) NKC labelled with indium-111 oxine with a tenfold excess of unlabelled cells during NKC therapy. Whole-body scintigrams were obtained (0.5-144 h) in the anterior and posterior views. Scintigrams were analysed using a region of interest technique, and single-photon emission tomography (SPET) studies of the abdomen were performed. Results were compared to those obtained with polymerase chain reaction (PCR) of the peripheral blood (determination of foreign DNA, nested PCR, limit of detection 0.01%). Shortly after transfusion of NKC, more than 50% of the activity was accumulated in the lungs. We observed redistribution effects from lungs to liver, spleen and bone marrow. No significant loss of activity could be detected. In two of four large metastases, tracer accumulation could be proven by SPET. As confirmed by scintigrams and PCR, the fraction of circulating transfused cells was low at all times. Long-term activity retention might be caused either by survival of the allogenic cells, as confirmed by PCR (up to 3 days p.i.), or by phagocytosis of labelled cellular fragments. However, PCR data and uptake in metastases indicated long survival of a portion of allogenic NKC. Such long survival and low retention of the cells in the lung are requirements for an effective immunotherapeutic approach.

The role of myoepithelial cells in the short-term radiogenic impairment of salivary glands. An immunohistochemical, ultrastructural and scintigraphic study.

BACKGROUND AND PURPOSE: The aim of this study was to evaluate the early dose-related functional impairment of salivary glands after radiation, using sialoscintigraphy, immunohistochemistry and electron microscopy in an established rabbit experimental model. MATERIALS AND METHODS: Twelve rabbits were used for the study. Eight were scintigraphically examined prior to and 24 hours after 15/30 Gy (4 rabbits each). The irradiated glands were examined immunohistochemically using monoclonal antibodies to alpha-smooth muscle actin (ASMA), vimentin and Ki-67 proliferation antigen. Ultrastructural investigation was also performed. Four control rabbits were sham-treated and provided normal salivary gland tissue. RESULTS: There was a significant increase of the 99mTc-pertechnetate uptake in the irradiated parotid glands (p < 0.05) and a highly significant one in the superficial mandibular glands (p < 0.001). Immunohistochemically a significant loss of ASMA and vimentin-stained myoepthelial cells and a decrease of the proliferating rate in the acinar cells could be assessed in both irradiated glands. Ultrastructurally, rarefaction and focal condensation of the myofilaments of the myoepithelial cells in both irradiated glands was observed. No dose-related effect could be assessed. CONCLUSION: Early functional impairment of salivary glands after radiation could be revealed as early as 24 hours after radiation. The considerable myoepthelial cell impairment may explain the secretory retention assessed scintigraphically and provide -apart from acinar cell damage--a new aspect in the pathogenesis of radiogenic impairment of salivary glands.