2.

4 Rheological measurements of polymer/mucus

mixtures
In order to evaluate the interactions between mucus and polymers, the dynamic viscosity of their mixtures were
measured. PAAs were dissolved and Carbopol 971P, Carbopol 974P, Carbopol 980, polycarbophil, Pemulen TR1, Pemulen TR-2, Ultrez 20, and Ultrez 21 were hydrated in 0.1 M phosphate buffer pH 6.8 (at concentration of 1
% w/v). Then, 1 mL of each polymer in buffer was mixed with 1 g of small intestinal porcine mucus and incubated
for 30 minutes at 37 C. Control without polymer was mixed with buffer and samples of those polymers in buffer
alone were also incubated at the same condition. Dynamic viscosity was measured in a cone-plate rheometer
(Haake Mars Rheometer, 379-0200, Thermo Electron GmbH, Karlsruhe, Germany), the oscillating measurements
were performed with a shear stress range of 0.510 Pa and the maintained temperature at 37 0.1 C.

2.5 Rotating cylinder method

Mucoadhesive properties of polymers were analyzed via rotating cylinder method [20] with 3 different porcine
mucosa (intestinal, vaginal, and buccal mucosa). Porcine mucosas were fixed on stainless steel cylinders and
tablets were attached to the mucosa gently. The cylinder was then used as stirrer in a dissolution tester (Erweka
GmbH, Heusenstamm, Germany) following to the European Pharmacopeia (Paddle Apparatus). The vessels were
filled with 900 mL of buffers with temperature maitained at 37 0.1C. The speed of cylinders were set at 125 rpm
and the detachment or complete disintegration of tablets was observed visually.
Different buffers were used to mimic biological media. Therefore, 0.1 M phosphate buffer pH 6.8 was used for
porcine intestinal mucosa experiments, simulated vaginal fluid pH 4.2 was used for porcine intestinal vaginal
mucosa experiments, and 0.1 M phosphate buffer pH 6.0 was used for porcine buccal mucosa experiments.
The simulated vaginal fluid contains sodium chloride 3.5 g/L, potassium hydroxide 1.4 g/L, calcium hydroxide 0.22
g/L, lactic acid 2.0 g/L, acetic acid 1.0 g/L, glycerol 0.16 g/L, urea 0.4 g/L, bovine serum albumin 0.018 g/L, and
glucose 5.0 g/L in demineralized water.

2.7 Tensile studies

Tensile studies were performed with 3 different porcine mucosa (intestinal, vaginal, and buccal mucosa). The
mucosa was cut into pieces of similar size of 2 2 (cm) and glued at the bottom of a beaker. One tablet was
glued to a 10-mm stainless steel flat disc which was hung from a laboratory stand with a nylon thread (15 cm).
The beaker was placed on a balance, and then slowly raised by height-adjustable stander until mucosa and tablet
came into contact. The beaker was filled with 100 mL buffer (specific buffer corresponding to type of mucosa used
as in section 2.3) and the setting was placed standstill for 5 minutes at room temperature. Afterward, the beaker
was moved down away from the hung tablet at a rate of 0.1 mm/s by by turning the knod of the height-adjustable
stander. Data were collected by computer software (SartoCollect V 1.0; Satorius AG, Germany) connected to the
balance with integrated interface. The force versus displacement curve was analysed to calculate the maximum
detachment force (MDF) and the total work of adhesion (TWA) as the area under the curve in accordance with the
trapezoidal rule. The experiment was repeated three times with each polymer and type of porcine mucosa.