INTRODUCTION:

Oral health is the mirror for the general individual health ¹. Many studies try to find the causes for the changes in the rate ofsalivary flow, changes in biochemical ingredients of saliva and the levels of minerals, immunoglobulins, enzymes electrolytes and proteins^2,3,4. One of the causes is the irradiation of the head and neck and even the total body irradiation. Radiotherapy is used to destroy tumors cells as an oncological treatment, in the same time normal tissues may damaged including mucus membrane, salivary glands , bone and even the teeth^5,6,7,8. Along with destruction of oral tissues, signs and symptoms are arise with alteration in biochemical components of the saliva^9,10,11,12.

Many studies showed changes in saliva composition due to sensitivity of salivary gland tissue to radiation^{13,14,15, 16}, therefore, the studies were recorded reduction in saliva volume, reduction in buffer capacity, more viscosity, acidic pH, changes in mucoglycoprotein, total protein and IgA levels that may persisted up to 4-6 months after the end of the treatment.

The current study was designed to evaluate the salivary flow rate, buffering capacity, amylase activity, and total protein levels in patients with radiation in comparison to cancer-free controls.

MATERIALS AND METHODS:

Subjects:

The participants of this study were composed of sixty human male volunteers (age 25 to 50 years), thirty subjects of them are patients withhead and neck cancer (experimental group) submitted to irradiation treatment at Al-Andalus oncology hospital‎in Baghdad while the other thirty group composed of healthy control subjects. All participants showed a good general health and were not taking any medication that capable to change the production of saliva and all of them were submitted to saliva collection.

Collection of the saliva:

Two ml of stimulated saliva were obtained after using a piece of Paraffin chewing film (USA). Collection of saliva related to group radiotherapy was done before and after radiation (received 35 daily, between 50 to 70 cGy) and between 8 -9 a.m. Other collection was done for control subjects. Salivary flow was recorded by measuring the time necessary (seconds) for the output of 2ml of stimulated saliva.

Prior one hour of saliva collection, all patients and control subjects were not eat, drink or smoke. The collected saliva was centrifuged at (16,000g) at 4ºC for 10 minutes and then kept in aliquot tubes and stored at (-70ºC) until biochemical evaluation.

Biochemical analysis:

Buffering capacity:

Evaluation of the salivary buffering capacity was done according to Spadaro et al 1998¹⁷. The pH of the saliva was estimated by using a simple colorimetric method a 0.5ml of the sample with 0.5ml of hydrochloric acid 12 mmol/l was analyzed with 0.05% of bromocresol green.

Protein determination:

The total protein content for the centrifuged samples 5 mg/sample was estimated by usingpyrogallol red dye by the endpoint method.Values were expressed as mg/dl.Martinezet al 2007¹⁸.

3. Salivary amylase activity:

Salivary amylase activity was measured at 25ºC with the dilution of saliva (250 times) in 0.05mol/l phosphate buffer at pH (6.9) and starch was used as a substrate. The reducing sugars that liberated by the action of enzyme were quantified by the di-nitro-salicylic acid method, using the glucose to plot the standard curve, ship 2000¹⁹.

Statistical analysis:

Statistical analysis of the data was performed by estimation of mean values ± standard deviation. And evaluate statistical significance between the study groups by unpaired Student’s t-test. The differences were evaluated significant at a level of p < 0.05, (*), p < 0.01 (**) and p < 0.001 (***).

RESULTS:

Results for salivary flow rate are estimated for the all subjects involved in the study,show significant increment for the experimental group after irradiation in comparison to control group (410±32.22; 160±27.12, respectively). While a non significant different value is found in comparison of experimental group before irradiation with control group. For buffer capacity, results show statistically significant reduction (p < 0.01) for the experimental group after irradiation in comparison to control group (pH 4.50±0.13; 5.33±0.14, respectively). No statistically significant alteration was observed in buffer capacity in comparison of experimental group before irradiation with control group, table (1). Statistically significant reduction of salivary a-amylase activity and total protein concentration are recorded in current results (130.22±20.01 before and 104.22±28.42 after irradiation in comparison to control) and (93.43±5.66 before and 87.54±6.13 after radiation in comparison to control), respectively, table (2).

Table 1: Salivary flow rate and buffering capacity for the experimental group (before and after radiotherapy) in comparison to the control group

Parameters	Total Sample No.	Study groups
		control	Experimental
		control	Before	After
Salivary flow rate (in seconds)	60	160 ± 27.12	170.32± 20.11	410± 32.22**
Buffer capacity	60	5.33± 0.14	5.47± 0.16	4.50± 0.13**

P< 0.01 (** )

Table 2: Salivary amylase (U/ml) and salivary total protein (mg/dl) for the experimental group (before and after radiotherapy) in comparison to the control group

Parameters	Total Sample No.	Study groups
		control	Experimental
		control	Before	After
Salivary amylase(U/ml)	60	155.96± 23.12	130.22± 20.01*	104.22± 28.42**
Total protein con. (mg/dl)	60	105.23±5.11	93.43± 5.66 *	87.54± 6.13***

P< 0.05, (*), P < 0.01 (**) and P < 0.001 (***).

DISCUSSION:

Patients with radiotherapy specifically with the treatment of head and neck cancer illustrated a hypofunction of the salivary glands and showed xerostomia with decrease in salivary flow²⁰. In current study, the saliva was collected from an irradiated patients with head and neck cancer (before and after radiation) and examined the salivary flow rate which was found an increment in rate of flow in patients after irradiation with decreased in pH from neutral to acidic, that resulted in lower buffer capacity in comparison to control group. These changes may related to effect of radiation on tissues causing a change in the salivary ionic composition with the loss of water from the content of saliva^21,22,23.

The current results also show a decrease in salivary amylase and total protein concentration in an irradiate subjects before and after radiation in comparison to control. These results may contributed to effect of radiation on physical properties of mucin which record an alteration and the loss of water from it, causing the fibers to be thinner and the network to become fragile and easily degrade resulting in reduction of total protein ^24,25,26.

This study suggests that the recorded parameters (saliva flow, buffering capacity , salivary amylase and salivary total protein) were modified during radiotherapy and showed alteration in their concentrations and values.

CONCLUSIONS:

The current data show changes in salivary parameters which give an idea to understand the alterations that happen in tissues due to radiation treatment and helps the clinician to evaluate and adapt the constituents of patients’ saliva and try to improve the dryness of patient’s mouth and support the patient’s health.

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Received on 27.02.2022 Modified on 15.04.2022

Research J. Pharm. and Tech 2023; 16(1):184-186.

DOI: 10.52711/0974-360X.2023.00034